EP4746902A2

EP4746902A2 - Compositions for delivery of plasmodium antigens and related methods

Info

Publication number: EP4746902A2
Application number: EP24754502.3A
Authority: EP
Inventors: Adam ZUIANI; Charles Lefco DULBERGER; Asaf PORAN; Daniel Abram Rothenberg; John SROUJI; Charles JENNISON; Patricia DOS SANTOS MEIRELES; Anja DOKIC; Annette VOGEL
Original assignee: BioNTech SE
Current assignee: BioNTech SE
Priority date: 2023-07-21
Filing date: 2024-07-19
Publication date: 2026-05-27
Also published as: AU2024301077A1; CN121816193A; JP2026524968A; WO2025024335A3; WO2025024335A2; IL325865A

Abstract

The present disclosure provides compositions (e.g., pharmaceutical compositions) for delivery of malarial protein antigens and related technologies (e.g., components thereof and/or methods relating thereto). Among other things, the present disclosure provides polyribonucleotides encoding malarial protein antigens.

Description

COMPOSITIONS FOR DELIVERY OF PLASMODIUM ANTIGENS AND RELATED METHODS

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to United States Provisional Application Serial No. 63/515,079, filed July 21, 2023, the entirety of which is incorporated herein by reference.

BACKG ROUND

[0002] Malaria is a mosquito-borne infectious disease caused by protozoan parasites of the Plasmodium genus.

According to the World Health Organization, an estimated 3.4 billion people in 92 countries are at risk of being infected with the malaria parasite and developing disease.

SUMMARY

[0003] The present disclosure provides technologies (e.g., compositions, methods, etc) for delivery of Plasmodium antigens (also referred to herein as "malaria antigens”). In one aspect, provided herein is a polyribonucleotide encoding a polypeptide, wherein the polypeptide comprises one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof. In some embodiments, a portion is an antigenic portion.

[0004] In some embodiments, a polyribonucleotide as provided herein is an isolated polyribonucleotide. In some embodiments, a polyribonucleotide as provided herein is an engineered polyribonucleotide. In some embodiments, a polyribonucleotide as provided herein is a codon-optimized polyribonucleotide.

[0005] In some embodiments, one or more Plasmodium Rh5 invasion complex antigens comprise: (i) one or more Plasmodium reticulocyte-binding protein homolog 5 (Rh5) polypeptides or antigenic portions thereof; (ii) one or more Plasmodium Cysteine-Rich Protective Antigen (CyRPA) polypeptides or antigenic portions thereof; (iii) one or more Plasmodium Rh5-interacting Protein (Ripr) polypeptides or antigenic portions thereof; (iv) one or more Plasmodium P113 polypeptides or antigenic portions thereof; (v) one or more Plasmodium thrombospondin-related apical merozoite protein (TRAMP) polypeptides or antigenic portions thereof; or (vi) one or more Plasmodium cysteine-rich small secreted protein (CSS) polypeptides or antigenic portions thereof.

[0006] In some embodiments, a polyribonucleotide encodes a polypeptide, wherein the polypeptide comprises one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof. In some embodiments, a polyribonucleotide encodes a polypeptide, wherein the polypeptide comprises one or more Plasmodium reticulocytebinding protein homolog 5 (Rh5) polypeptides or antigenic portions thereof.

[0007] In some embodiments, one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof comprise one or more antigenic portions of Plasmodium Rh5. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise one or more ordered domains of Rh5. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise two ordered domains of Rh5. In some embodiments, two ordered domains of Rh5 are directly adjacent to one another. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise a cysteine at position 203, 329, or both, as numbered according to SEQ ID NO: 1. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise: (i) an amino acid sequence according to SEQ ID NO: 118, 119, 124, or 126, (ii) an amino acid sequence according to SEQ ID NO: 120 or 128, or (iii) a combination thereof.

RECTIFIED SHEET (RULE 91) ISA/EP [0008] In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise a tyrosine at position 203, 329, or both, as numbered according to SEQ ID NO: 1. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise: (i) an amino acid sequence according to SEQ ID NO: 121, 122, 125, or 127, (ii) an amino add sequence according to SEQ ID NO: 123 or 129, or (iii) a combination thereof. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise a tyrosine at position 203, as numbered according to SEQ ID NO: 1. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise a tyrosine at position 329, as numbered according to SEQ ID NO: 1.

[0009] In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise one, two, three, or four N-linked glycosylation sites. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise two N-llnked glycosylation sites. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise an amino acid substitution at one or more N-linked glycosylation sites, wherein the amino acid substitution prevents glycosylation. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise a substitution of NX[T/S] to QX[T/S] and/or a substitution of NX[T/S] to NXA. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise: (i) an amino add sequence according to SEQ ID NO: 124, 125, 126, or 127, (ii) an amino add sequence according to SEQ ID NO: 128 or 129, or (ii) a combination thereof.

[0010] In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise an amino acid substitution that prevents glycosylation at position 214, as numbered according to SEQ ID NO: 1. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise an amino acid substitution that prevents glycosylation at position 297, as numbered according to SEQ ID NO: 1. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise an amino add substitution at all of the N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation.

[0011] In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise a PMX deavage site. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise an amino add sequence according to SEQ ID NO: 118, 121, 124, or 125. In some embodiments, one or more Plasmodium Rh5 antigenic portions do not comprise a PMX cleavage site.

[0012] In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise an amino add sequence according to SEQ ID NO: 119, 122, 126, or 127. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise: (i) an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 130, (ii) an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 118, 119, 121, 122, 124, 125, 126, or 127, (iii) an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 131, (iv) an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 120, 123, 128, or 129, or (v) a combination thereof.

[0013] In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise (i) an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 118, 119, 121, 122, 124, 125, 126, or 127, (ii) an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 120, 123, 128, or 129, or (iii) a combination thereof. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise: (i) an amino add sequence according to SEQ ID NO: 121 or 122, and (ii) an amino add sequence according to SEQ ID NO: 120. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise: (i) an amino acid sequence according to SEQ ID NO: 125 or 127, and (ii) an amino add sequence according to SEQ ID NO: 120 or 128.

[0014] In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise an amino acid sequence with at least 85% sequence identity to an amino add sequence of positions 26-363 of SEQ ID NO: 94, 95 or 99. In some embodiments, one or more PlasmotSum Rh5 antigenic portions comprise an amino acid sequence with at least 85% sequence identity to an amino add sequence of positions 26-526 of SEQ ID NO: 89. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise an amino add sequence of positions 26-363 of SEQ ID NO: 94. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise an amino add sequence of positions 26-363 of SEQ ID NO: 95. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise an amino acid sequence of positions 26-363 of SEQ ID NO: 99. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise an amino add sequence of positions 26-526 of SEQ ID NO: 89. In some embodiments, one or more Plasmodium Rh5 antigenic portions do not comprise a disordered domain of Rh5. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise a PMX cleavage site. In some embodiments, a PMX deavage site comprises or consists of an amino acid sequence of NFLQ.

[0015] In some embodiments, one or more Plasmodium Rh5 Invasion complex antigens comprise a Plasmodium CyRPA polypeptide or antigenic portion thereof. In some embodiments, one or more Plasmodium Rh5 Invasion complex antigens comprise an antigenic portion of Piasmocfium CyRPA.

[0016] In some embodiments, a Plasmodium CyRPA antigenic portion comprises an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 133. In some embodiments, a Plasmodium CyRPA antigenic portion comprises or consists of an amino add sequence of SEQ ID NO: 133. In some embodiments, a Plasmodium CyRPA antigenic portion comprises an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 169. In some embodiments, a Plasmodium antigenic portion comprises or consists of an amino acid sequence of SEQ ID NO: 169.

[0017] In some embodiments, a Plasmodium CyRPA antigenic portion comprises one, two, or three N-llnked glycosylation sites. In some embodiments, a Plasmodium CyRPA antigenic portion comprises an amino add substitution at one or more N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation. In some embodiments, a Plasmodium CyRPA antigenic portion comprises a substitution of NX[T/S] to QX[T/S]. In some embodiments, a Plasmodium CyRPA antigenic portion comprises a substitution of NX[T/S] to NXA. In some embodiments, a Plasmodium CyRPA antigenic portion comprises an amino add substitution at all of the N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation.

[0018] In some embodiments, a Plasmodium CyRPA antigenic portion comprises an asparagine at position 145, position 322, position 338, or a combination thereof, as numbered according to SEQ ID NO: 3. In some embodiments, a Plasmodium CyRPA antigenic portion comprises a glutamine at position 145, position 322, position 338, or a combination thereof, as numbered according to SEQ ID NO: 3. In some embodiments, a Plasmodium CyRPA antigenic portion comprises a glutamine at position 145, position 322, and position 338, as numbered according to SEQ ID NO: 3. In some embodiments, a Plasmodium CyRPA antigenic portion comprises or consists of an amino acid sequence of SEQ ID NO: 134. In some embodiments, a Plasmodium CyRPA antigenic portion comprises or consists of an amino add sequence of SEQ ID NO: 170. [0019] In some embodiments, one or more Plasmodium Rh5 invasion complex antigens comprise a Plasmodium Pl 13 polypeptide or antigenic portion thereof. In some embodiments, one or more Plasmodium Rh5 invasion complex antigens comprise an antigenic portion of Plasmodium Pl 13.

[0020] In some embodiments, a Plasmodium P113 antigenic portion comprises an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 135. In some embodiments, a Plasmodium P113 antigenic portion comprises or consists of an amino acid sequence of SEQ ID NO: 135.

[0021] In some embodiments, a Plasmodium P113 antigenic portion comprises one, two, three, four, five, six, seven, or eight N-linked glycosylation sites. In some embodiments, a Plasmodium P113 antigenic portion comprises an amino acid substitution at one or more N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation. In some embodiments, a Plasmodium P113 antigenic portion comprises a substitution of NX[T/S] to QX[T/S], In some embodiments, a Plasmodium P113 antigenic portion comprises a substitution of NX[T/S] to NXA. In some embodiments, a Plasmodium P113 antigenic portion comprises an amino add substitution at all of the N-linked glycosylation sites, wherein the amino acid substitution prevents glycosylation.

[0022] In some embodiments, a Plasmodium P113 antigenic portion comprises an asparagine at position 207, position 268, position 317, position 360, position 661, position 697, position 779, position 876, or a combination thereof, as numbered according to SEQ ID NO: 6. In some embodiments, a Plasmodium Pl 13 antigenic portion comprises a glutamine at position 207, position 268, position 317, position 360, position 661, position 697, position 779, position 876, or a combination thereof, as numbered according to SEQ ID NO: 6. In some embodiments, a Plasmodium Pl 13 antigenic portion comprises a glutamine position 207, position 268, position 317, position 360, position 661, position 697, position 779, and position 876, as numbered according to SEQ ID NO: 6. In some embodiments, a Plasmodium P113 antigenic portion comprises or consists of an amino add sequence of SEQ ID NO: 136.

[0023] In some embodiments, one or more Plasmodium Rh5 invasion complex antigens comprise a Plasmodium Rlpr polypeptide or antigenic portion thereof. In some embodiments, one or more Plasmodium Rh5 invasion complex antigens comprise an antigenic portion of Plasmodium Rlpr.

[0024] In some embodiments, a Plasmodium Rlpr antigenic portion comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 178. In some embodiments, a Plasmodium Ripr antigenic portion comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 177. In some embedments, a Plasmodium Ripr antigenic portion comprises or consists of an amino add sequence according to SEQ ID NO: 177. [0025] In some embodiments, a Plasmodium Ripr antigenic portion comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 176. In some embodiments, a Plasmodium Ripr antigenic portion comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 175. In some embodiments, a Plasmodium Ripr antigenic portion comprises or consists of an amino add sequence according to SEQ ID NO: 175. [0026] In some embodiments, a Plasmodium Ripr antigenic portion comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 174. In some embodiments, a Plasmodium Ripr antigenic portion comprises or consists of an amino acid sequence according to SEQ ID NO: 174. [0027] In some embodiments, a Plasmodium Ripr antigenic portion comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 190. In some embodiments, a Plasmodium Ripr antigenic portion comprises or consists of an amino acid sequence according to SEQ ID NO: 190.

[0028] In some embodiments, a Plasmodium Ripr antigenic portion comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 171. In some embodiments, a Plasmodium Ripr antigenic portion comprises or consists of an amino acid sequence according to SEQ ID NO: 171.

[0029] In some embodiments, a Plasmodium Ripr antigenic portion comprises one, two, three, four, five, six, seven, eight, nine, ten, eleven or twelve N-llnked glycosylation sites. In some embodiments, a Plasmodium Ripr antigenic portion comprises an amino acid substitution at one or more N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation. In some embodiments, a Plasmodium Ripr antigenic portion comprises a substitution of NX[T/S] to QX[T/S]. In some embodiments, a Plasmodium Ripr antigenic portion comprises an amino add substitution at all of the N-linked glycosylation sites, wherein the amino acid substitution prevents glycosylation. [0030] In some embodiments, a Plasmodium Ripr antigenic portion comprises an asparagine at position 646, position 964, position 1021, or a combination thereof, as numbered according to SEQ ID NO: 2. In some embodiments, a Plasmodium Ripr antigenic portion comprises a glutamine at position 646, position 964, position 1021, or a combination thereof, as numbered according to SEQ ID NO: 2. In some embodiments, a Plasmodium Ripr antigenic portion comprises a glutamine at position 646, position 964, and position 1021, as numbered according to SEQ ID NO: 2. In some embodiments, a Plasmodium Ripr antigenic portion comprises or consists of an amino add sequence according to SEQ ID NO: 190.

[0031] In some embodiments, a Plasmodium Ripr antigenic portion comprises an asparagine at position 103, position 144, position 228, position 303, position 334, position 480, position 498, position 506, position 526, position 646, position 964, position 1021, or a combination thereof, as numbered according to SEQ ID NO: 2. In some embodiments, a Plasmodium Ripr antigenic portion comprises a glutamine at position 103, position 144, position 228, position 303, position 334, position 480, position 498, position 506, position 526, position 646, position 964, position 1021, or a combination thereof, as numbered according to SEQ ID NO: 2. In some embodiments, a Plasmodium Ripr antigenic portion comprises a glutamine at position 103, position 144, position 228, position 303, position 334, position 480, position 498, position 506, position 526, position 646, position 964, position 1021, as numbered according to SEQ ID NO: 2. In some embodiments, a Plasmodium Ripr antigenic portion comprises or consists of an amino add sequence according to SEQ ID NO: 171.

[0032] In some embodiments, a Plasmodium Ripr antigenic portion comprises a PMX cleavage site. In some embodiments, a PMX deavage site comprises or consists of an amino acid sequence of SMLE.

[0033] In some embodiments, one or more Plasmodium Rh5 invasion complex antigens comprise a Plasmodium TRAMP polypeptide or antigenic portion thereof. In some embodiments, one or more Plasmodium Rh5 invasion complex antigens comprise an antigenic portion of Plasmodium TRAMP.

[0034] In some embodiments, a Plasmodium TRAMP antigenic portion comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 179. In some embodiments, a PfasmodiumTRMAP antigenic portion comprises or consists of an amino add sequence according to SEQ ID NO: 179. [0035] In some embodiments, a Plasmodium TRAMP antigenic portion comprises one, two, three, four, five, six, seven, or eight N-linked glycosylation sites. In some embodiments, a Plasmodium TRAMP antigenic portion comprises an amino acid substitution at one or more N-linked glycosylation sites, wherein the amino acid substitution prevents glycosylation. In some embodiments, a Plasmodium TRAMP antigenic portion comprises a substitution of NX[T/S] to QX[T/S], In some embodiments, a /¥asmoa6i/fl)TRAMP antigenic portion comprises an amino add substitution at all of the N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation. [0036] In some embodiments, a PiasmodmmTRMAP antigenic portion comprises an asparagine at position 149, position 195, position 202, or a combination thereof, as numbered according to SEQ ID NO: 4. In some embodiments, a Plasmodium TRAMP antigenic portion comprises a glutamine at position 149, position 195, position 202, or a combination thereof, as numbered according to SEQ ID NO: 4. In some embodiments, a Plasmodium TRAMP antigenic portion comprises a glutamine at position 149, position 195, and position 202, as numbered according to SEQ ID NO: 4. In some embodiments, a Plasmodium TRAMP antigenic portion comprises or consists of an amino add sequence according to SEQ ID NO: 181.

[0037] In some embodiments, a Plasmodium TRAMP antigenic portion comprises an asparagine at position 112, position 149, position 155, position 170, position 195, position 202, position 253, position 305, or a combination thereof, as numbered according to SEQ ID NO: 4. In some embodiments, a Plasmodium TRAMP antigenic portion comprises a glutamine at position 112, position 149, position 155, position 170, position 195, position 202, position 253, position 305, or a combination thereof, as numbered according to SEQ ID NO: 4. In some embodiments, a Plasmodium TRAMP antigenic portion comprises a glutamine at position 112, position 149, position 155, position 170, position 195, position 202, position 253, and position 305, as numbered according to SEQ ID NO: 4. In some embodiments, a Plasmodium TRAMP antigenic portion comprises or consists of an amino acid sequence according to SEQ ID NO: 180.

[0038] In some embodiments, a Plasmodium TRAMP antigenic portion comprises a PMX deavage site. In some embodiments, a PMX deavage site comprises or consists of an amino add sequence of HFLQ. In some embodiments, a Plasmodium TRAMP antigenic portion comprises a SUB2 cleavage site. In some embodiments, a SUB2 deavage site comprises or consists of an amino add sequence according to SEQ ID NO: 193.

[0039] In some embodiments, one or more Plasmodium Rh5 invasion complex antigens comprise a Ptasmodum CSS polypeptide or antigenic portion thereof. In some embodiments, one or more Plasmodium Rh5 invasion complex antigens comprise an antigenic portion of Plasmodium CSS.

[0040] In some embodiments, a Plasmodium CSS antigenic portion comprises one or more cysteine to serine mutations. In some embodiments, a Ptasmodum CSS antigenic portion comprises a C30S mutation, a C80S mutation, a C276S mutation, or a combination thereof. In some embodiments, a Plasmodium CSS antigenic portion comprises a serine at position 30 and position 80, as numbered according to SEQ ID NO: 5 or 214.

[0041] In some embodiments, a Plasmodium CSS antigenic portion comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 194. In some embodiments, a Plasmodium CSS antigenic portion comprises or consists of an amino add sequence according to SEQ ID NO: 194. In some embodiments, a Plasmodium CSS antigenic portion comprises a serine at position 30 and position 276, as numbered according to SEQ ID NO: 5 or 214.

[0042] In some embodiments, a Plasmodium CSS antigenic portion comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 195. In some embodiments, a Plasmodium CSS antigenic portion comprises or consists of an amino acid sequence according to SEQ ID NO: 195.

[0043] In some embodiments, a Plasmodium CSS antigenic portion comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 182. In some embodiments, a Plasmodium CSS antigenic portion comprises or consists of an amino acid sequence according to SEQ ID NO: 182.

[0044] In some embodiments, a Plasmodium CSS antigenic portion comprises one, two, three, four, five, or six N-inked glycosylation sites. In some embodiments, a Plasmodium CSS antigenic portion comprises an amino acid substitution at one or more N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation. In some embodiments, a Plasmodium CSS antigenic portion comprises a substitution of NX[T/S] to QX[T/S], In some embodiments, a Plasmodium CSS antigenic portion comprises an amino add substitution at all of the N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation.

[0045] In some embodiments, a Plasmodium CSS antigenic portion comprises an asparagine at position 74, position 88, or a combination thereof, as numbered according to SEQ ID NO: 5 or 214. In some embodiments, a Plasmodium CSS antigenic portion comprises a glutamine at position 74, position 88, or a combination thereof, as numbered according to SEQ ID NO: 214.

[0046] In some embodiments, a Plasmodium CSS antigenic portion comprises a glutamine at position 74 and position 88, as numbered according to SEQ ID NO: 5 or 214. In some embodiments, a Plasmodium CSS antigenic portion comprises or consists of an amino add sequence according to SEQ ID NO: 184.

[0047] In some embodiments, a Plasmodium CSS antigenic portion comprises an asparagine at position 192, position 234, position 261, position 283, or a combination thereof, as numbered according to SEQ ID NO: 195. In some embodiments, a Plasmodium CSS antigenic portion comprises a glutamine at position 192, position 234, position 261, position 283, or a combination thereof, as numbered according to SEQ ID NO: 195. In some embodiments, a Plasmodium CSS antigenic portion comprises a glutamine at position 192, position 234, position 261, and position 283, as numbered according to SEQ ID NO: 195. In some embodiments, a Plasmodium CSS antigenic portion comprises or consists of an amino add sequence according to SEQ ID NO: 185.

[0048] In some embodiments, a Plasmodium CSS antigenic portion comprises an asparagine at position 74, position 88, position 192, position 234, position 261, position 283, or a combination thereof, as numbered according to SEQ ID NO: 182. In some embodiments, a Plasmodium CSS antigenic portion comprises a glutamine at position 74, position 88, position 192, position 234, position 261, position 283, or a combination thereof, as numbered according to SEQ ID NO: 182. In some embodiments, a Plasmodium CSS antigenic portion comprises a glutamine at position 74, position 88, position 192, position 234, position 261, and position 283, as numbered according to SEQ ID NO: 182. In some embodiments, a Plasmodium CSS antigenic portion comprises or consists of an amino add sequence according to SEQ ID NO: 183.

[0049] In some embodiments, a polypeptide comprises two or more Piasmodum Rh5 invasion complex polypeptides or antigenic portions thereof. In some embodiments, Rh5 invasion complex polypeptides or antigenic portions thereof comprise two selected from (I) one or more Plasmodium Rh5 polypeptides or antigenic portions thereof, (ii) one or more Plasmodium C'fRPk polypeptides or antigenic portions thereof, (iii) one or more Plasmodium Ripr polypeptides or antigenic portions thereof, (iv) one or more Plasmodium Pl 13 polypeptides or antigenic portions thereof, (v) one or more Plasmodium TRAMP polypeptides or antigenic portions thereof, and (vi) one or more Plasmodium CSS polypeptides or antigenic portions thereof.

[0050] In some embodiments, a polypeptide comprises one or more Plasmodium Rh5 antigenic portions and further comprises: (i) one or more Plasmodium CyRPA polypeptides or antigenic portions thereof, (ii) one or more Plasmodium Ripr polypeptides or antigenic portions thereof, (iii) one or more Plasmodium P113 polypeptides or antigenic portions thereof, (iv) one or more Ptis/nodt/m TRAMP polypeptides or antigenic portions thereof, or (v) one or more Plasmodium CSS polypeptides or antigenic portions thereof.

[0051] In some embodiments, a polypeptide comprises one or more Plasmodium Rh5 antigenic portions, wherein the polypeptide comprises: (i) one or more Plasmodium Rh5 polypeptides or antigenic portions thereof and one or more Plasmodium CyRPA polypeptides or antigenic portions thereof; (il) one or more Plasmodium Rh5 polypeptides or antigenic portions thereof and one or more Plasmodium Pl 13 polypeptides or antigenic portions thereof; (iii) one or more Plasmodium Rh5 polypeptides or antigenic portions thereof and one or more a Plasmodium Ripr polypeptides or antigenic portions thereof; (hr) one or more Plasmodium Rh5 polypeptides or antigenic portions thereof and one or more a Plasmodium TRAMP polypeptides or antigenic portions thereof; or (v) one or more Plasmodium Rh5 polypeptides or antigenic portions thereof and one or more a Plasmodium CSS polypeptides or antigenic portions thereof.

[0052] In some embodiments, a polypeptide comprises a secretory signal. In some embodiments, a secretory slgial is located at the N-terminus of the polypeptide.

[0053] In some embodiments, a secretory signal is a heterologous secretory signal. In some embodiments, a heterologous secretory signal comprises or consists of a viral secretory signal. In some embodiments, a viral secretory signal comprises or consists of an HSV secretory signal. In some embodiments, an HSV secretory signal comprises or consists of an HSV-1 or HSV-2 secretory signal. In some embodiments, an HSV secretory signal comprises or consists of an HSV glycoprotein D (gD) secretory signal. In some embodiments, an HSV gD secretory signal consists of an amino add sequence according to SEQ ID NO: 42.

[0054] In some embodiments, a secretory signal comprises or consists of a Plasmodium seaetarf signal. In some embodiments, a secretory signal comprises or consists of a Plasmodium CyRPA secretory signal. In some embodiments, a secretory signal comprises or consists of a Plasmodium Ripr secretory signal. In some embodiments, a secretory signal comprises or consists of a Plasmodium CSS secretory signal. In some embodiments, a secretory signal comprises or consists of an amino add sequence according to SEQ ID NOs: 186, 187, 188.

[0055] In some embodiments, a polypeptide comprises a transmembrane region. In some embodiments, a transmembrane region comprises a heterologous transmembrane region.

[0056] In some embodiments, a transmembrane region comprises or consists of a Plasmodium transmembrane region. In some embodiments, a transmembrane region comprises or consists of a P113 Plasmodium transmembrane region. In some embodiments, a transmembrane region comprises or consists of a TRAMP Plasmodium transmembrane region.

[0057] In some embodiments, a heterologous transmembrane region comprises or consists of a viral transmembrane region. In some embodiments, a heterologous transmembrane region comprises or consists of an HSV transmembrane region. In some embodiments, an HSV transmembrane region comprises or consists of an HSV- 1 or HSV-2 transmembrane region. In some embodiments, an HSV transmembrane region comprises or consists of an HSV gD transmembrane region. In some embodiments, an HSV gD transmembrane region consists of an amino add sequence according to SEQ ID NO: 75.

[0058] In some embodiments, a polypeptide does not comprise a transmembrane region. [0059] In some embodiments, a polypeptide comprises a multimerization domain. In some embodiments, a multimerization domain is a trimerization domain. In some embodiments, a trimerization domain is a C -terminal domain of T4 fibritin (e.g., foldon domain). In some embodiments, a foldon domain comprises or consists of an amino acid sequence according to SEQ ID NO: 78.

[0060] In some embodiments, a polypeptide comprises a self-assembling nanoparticle domain. In some embodiments, a self-assembling nanopartide domain is a ferritin domain. In some embodiments, a ferritin domain is from H. pylori. In some embodiments, a ferritin domain comprises or consists of a sequence according to SEQ ID NO. 88.

[0061] In some embodiments, a polypeptide comprises one or more linkers. In some embodiments, a polypeptide comprises one or more glydne-serine linkers. In some embodiments, one or more linkers comprise or consist of an amino add sequence according to SEQ ID NO: 80. In some embodiments, one or more linkers comprise or consist of an amino add sequence according to SEQ ID NO: 83. In some embodiments, one or more linkers comprise or consist of an amino acid sequence according to SEQ ID NO: 84. In some embodiments, one or more linkers comprise or consist of an amino add sequence according to SEQ ID NO: 86. In some embodiments, one or more linkers comprise or consist of an amino acid sequence according to SEQ ID NO: 137. In some embodiments, one or more linkers comprise or consist of an amino add sequence according to SEQ ID NO: 138.

[0062] In some embodiments, a polypeptide comprises a linker after an amino add sequence of the one or more Plasmodium Rh5 Invasion complex polypeptides or antigenic portions thereof.

[0063] In some embodiments, Plasmodium Is Plasmodium faidparum. In some embodiments, Plasmodium falciparum is Plasmodium faidparum isolate 3D7.

[0064] In some embodiments, one or more Plasmodium Rh5 Invasion complex polypeptides or antigenic portions thereof are one or more P. faidparum Rh5 invasion complex polypeptides or antigenic portions thereof. In some embodiments, one or more Plasmodium Rh5 polypeptides or antigenic portions thereof are one or more P. falciparum Rh5 antigenic portions. In some embodiments, one or more Plasmodium Rh5 Invasion complex polypeptides or antigenic portions thereof are a P. falciparum CyRPA polypeptide or antigenic portion thereof. In some embodiments, one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof are a P. faidparum Pl 13 polypeptide or antigenic portion thereof. In some embodiments, one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof are a P. falciparum Ripr polypeptide or antigenic portion thereof. In some embodiments, one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof are a P. fakiparum TRAMP polypeptide or antigenic portion thereof. In some embodiments, one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof are a P. fakiparum CSS polypeptide or antigenic portion thereof.

[0065] Provide herein is a polyribonucleotide encoding a polypeptide, wherein the polypeptide comprises a secretory signal, and two Plasmodium Rh5 ordered domains.

[0066] In some embodiments, a polypeptide comprises (i) a secretory signal, (II) two Plasmodium Rh5 ordered domains, (iH) a linker, and (iv) a multimerization domain. In some embodiments, a polypeptide comprises (I) a secretory signal, (II) two Plasmodium Rh5 ordered domains, (ill) a linker, and (iv) a transmembrane region. In some embodiments, a polypeptide comprises (i) a secretory signal, (ii) two Plasmodium Rh5 ordered domains, (iii) a linker, and (iv) a self-assembling nanoparticle domain.

[0067] In some embodiments, a secretory signal is a viral secretory signal. In some embodiments, a secretory skyial is an HSV glycoprotein D (gD) secretory signal, optionally wherein the HSV gD secretory signal consists of an amino acid sequence according to SEQ ID NO: 42.

[0068] In some embodiments, a polypeptide does not comprise a Plasmodium Rh5 disordered domain. In some embodiments, two Plasmodium Rh5 ordered domains are directly adjacent.

[0069] In some embodiments, a polypeptide comprises a C203Y substitution, as numbered according to SEQ

ID NO: 1.

[0070] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 94. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 94.

[0071] In some embodiments, a linker comprises or consists of a glydne-serine linker. In some embodiments, a linker comprises or consists of an amino acid sequence according to SEQ ID NO: 86.

[0072] In some embodiments, a multimerization domain Is a C-terminal domain of T4 fibrttin (e.g., fokion domain). In some embodiments, a foldon domain comprises or consists of an amino acid sequence according to SEQ ID NO: 78.

[0073] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 90. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 90.

[0074] In some embodiments, a transmembrane region is a heterologous transmembrane region. In some embodiments, a transmembrane region is a viral transmembrane region. In some embodiments, a viral transmembrane region Is a HSV gD transmembrane region. In some embodiments, a HSV gD transmembrane region comprises or consists of an amino add sequence according to SEQ ID NO: 75.

[0075] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 92. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 92.

[0076] In some embodiments, a linker comprises or consists of a glycine-serine linker. In some embodiments, a linker comprises or consists of an amino acid sequence according to SEQ ID NO: 137.

[0077] In some embodiments, a self-assembling nanoparticle domain is a ferritin domain. In some embodiments, a ferritin domain comprises or consists of a sequence according to SEQ ID NO: 88.

[0078] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 96. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 96.

[0079] In some embodiments, Plasmodium Rh5 ordered domains comprise one or more N-linked glycosylation sites.

[0080] In some embodiments, Plasmodium Rh5 ordered domains comprise one NX[T/S] to QX[T/S] substitution, optionally wherein the substitution is a N214Q substitution, as numbered according to SEQ ID NO: 1. [0081] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 99. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 99.

[0082] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 98. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 98.

[0083] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 101. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 101.

[0084] In some embodiments, a polypeptide comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 100. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 100.

[0085] In some embodiments, Plasmodium Rh5 ordered domains comprise two NX[T/S] to QX[T/S] substitutions, optionally wherein the two substitutions comprise N214Q and N297Q substitutions. In some embodiments, Plasmodium Rh5 ordered domains comprise an amino add substitution at al of the N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation.

[0086] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 95. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 95.

[0087] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 91. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 91.

[0088] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 93. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 93.

[0089] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 97. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 97.

[0090] Also provided herein is a polyribonucleotide encoding a polypeptide, wherein the polypeptide comprises (i) a secretory signal, (ii) a Plasmodium Rh5 N-terminal disordered domain, (iii) a Plasmodium Rh5 ordered domain, (iv) a Plasmodium Rh5 linking disordered domain, and (v) a Plasmodium Rh5 C-terminal ordered domain.

[0091] In some embodiments, a secretory signal Is a heterologous secretory signal. In some embodiments, a heterologous secretory signal is a viral secretory signal. In some embodiments, a viral secretor signal is an HSV glycoprotein D (gD) secretory signal. In some embodiments, an HSV gD secretory signal comprises or consists of an amino acid sequence according to SEQ ID NO: 42.

[0092] In some embodiments, a polypeptide comprises one or more N-linked glycosylation sites. In some embodiments, a polypeptide comprises an amino acid substitution at one or more of the N-linked glycosylation sites, wherein the amino acid substitution prevents glycosylation, optionally wherein the polypeptide comprises an amino add substitution at all of the N-linked glycosylation sites. [0093] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 89. In some embodiments, a polypeptide is or comprises an amino add sequence according to SEQ ID NO: 89.

[0094] The present disdosure further provides a polyribonucleotide encoding a polypeptide, wherein the polypeptide comprises (i) a secretory signal, and (ii) a Plasmodium CyRPA polypeptide or antigenic portion thereof. [0095] In some embodiments, a polypeptide comprises (i) a secretory signal, (ii) a Plasmodium CyRPA polypeptide or antigenic portion thereof, (Si) a linker, and (iv) a multimerization domain.

[0096] In some embodiments, a polypeptide comprises (i) a secretory signal, (ii) a Plasmodium CyRPA polypeptide or antigenic portion thereof, and (iii) a transmembrane region.

[0097] In some embodiments, a polypeptide comprises (i) a seaetory signal, (II) a Plasmodium Of'RPk polypeptide or antigenic portion thereof, (iii) a linker, and (iv) a transmembrane region.

[0098] In some embodiments, a secretory signal is a heterologous secretory signal. In some embodiments, a heterologous secretory signal is a viral secretory signal. In some embodiments, a viral secretor signal is an HSV glycoprotein D (gD) secretory signal. In some embodiments, an HSV gD secretory signal comprises or consists of an amino acid sequence according to SEQ ID NO: 42.

[0099] In some embodiments, a secretory signal is a Plasmodium seatiorf signal. In some embodiments, a Plasmodium secretory signal Is a Plasmodium CyRPA seaetory signal. In some embodiments, a Plasmodium CyRPA secretory signal comprises or consists of an amino add sequence according to SEQ ID NO: 187. In some embodiments, a Plasmodium CyRPA polypeptide or antigenic portion thereof comprises or consists of an amino add sequence with at least 85% sequence Identity to a sequence comprising SEQ ID NO: 133.

[0100] In some embodiments, a Plasmodium CyRPA polypeptide or antigenic portion thereof comprises or consists of SEQ ID NO: 169 or 133. In some embodiments, a Plasmodium CyRPA polypeptide or antigenic portion thereof comprises or consists of amino adds 29-362 of SEQ ID NO: 169 or 133.

[0101] In some embodiments, a Plasmodium CyRPA polypeptide or antigenic portion thereof comprises one or more N-linked glycosylation sites. In some embodiments, a Plasmodium CyRPA polypeptide or antigenic portion thereof comprises an amino acid substitution at one or more of the N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation.

[0102] In some embodiments, a Plasmodium CyRPA polypeptide or antigenic portion thereof comprises a substitution of NX[T/S] to QX[T/S] and/or a substitution of NX[T/S] to NXA. In some embodiments, a Plasmodium CyRPA polypeptide or antigenic portion thereof comprises an N145Q, N322Q, N338Q or a combination thereof. In some embodiments, a Plasmodium CyRPA polypeptide or antigenic portion thereof comprises an N145Q, N322Q, N338Q.

[0103] In some embodiments, a linker comprises a glydne-serine linker. In some embodiments, a linker comprises or consists of an amino acid sequence according to SEQ ID NO: 86. In some embodiments, a linker comprises or consists of an amino add sequence according to SEQ ID NO: 138.

[0104] In some embodiments, a multimerization domain is a C-terminal domain of T4 fibritin (foldon domain). In some embodiments, a foldon domain comprises or consists of an amino add sequence according to SEQ ID NO: 78.

[0105] In some embodiments, a transmembrane region is a heterologous transmembrane region. In some embodiments, a transmembrane region is a viral transmembrane region. In some embodiments, a viral transmembrane region is a HSV gD transmembrane region. In some embodiments, a HSV gD transmembrane region comprises or consists of an amino acid sequence according to SEQ ID NO: 75.

[0106] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 140. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 140.

[0107] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 141. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 141.

[0108] In some embodiments, a polypeptide comprises or consists of an amino acid sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 142. In some embodiments, a polypeptide comprises or consists of an amino acid sequence according to SEQ ID NO: 142.

[0109] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 143. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 143.

[0110] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 144. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 144.

[0111] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 198. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 198.

[0112] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 199. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 199.

[0113] Further provided is a polyribonucleotide encoring a polypeptide, wherein the polypeptide comprises (I) a secretory signal, (H) a Plasmodium P113 polypeptide or antigenic portion thereof, (ill) a linker, and (Iv) a transmembrane region. In some embodiments, a polypeptide comprises: (I) a secretory signal, (li) a Plasmodium P113 polypeptide or antigenic portion thereof, (Hi) a linker, and (iv) a transmembrane region.

[0114] In some embodiments, a secretory signal is a heterologous secretory signal. In some embodiments, a heterologous secretory signal is a viral secretory signal. In some embodiments, a viral secretor signal is an HSV glycoprotein D (gD) secretory signal. In some embodiments, an HSV gD secretory signal comprises or consists of an amino acid sequence according to SEQ ID NO: 42.

[0115] In some embodiments, a Ifriker comprises a glydne-serine linker. In some embodiments, a linker comprises or consists of an amino add sequence according to SEQ ID NO: 86.

[0116] In some embodiments, a transmembrane region is a heterologous transmembrane region. In some embodiments, a transmembrane region is a viral transmembrane region. In some embodiments, a viral transmembrane region is a HSV gD transmembrane region. In some embodiments, a HSV gD transmembrane region comprises or consists of an amino add sequence according to SEQ ID NO: 75.

[0117] In some embodiments, a Plasmodium P113 polypeptide or antigenic portion thereof comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 135. In some embodiments, a Plasmodium P113 polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 135.

[0118] In some embodiments, a Plasmodium P113 polypeptide or antigenic portion thereof comprises one or more N-linked glycosylation sites. In some embodiments, a Plasmodium P113 polypeptide or antigenic portion thereof comprises an amino acid substitution at one or more of the N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation. In some embodiments, a Plasmodium P113 polypeptide or antigenic portion thereof comprises one or more substitutions of NX[T/S] to QX[T/S] and/or substitutions of NX[T/S] to NXA.

[0119] In some embodiments, a Plasmodium P113 antigenic portion comprises an asparagine at position 207, position 268, position 317, position 360, position 661, position 694, position 779, position 876, or a combination thereof, as numbered according to SEQ ID NO: 6. In some embodiments, a Plasmodium P113 antigenic portion comprises a glutamine at position 207, position 268, position 317, position 360, position 661, position 694, position 779, position 876, or a combination thereof, as numbered according to SEQ ID NO: 6. In some embodiments, a Plasmodium P113 antigenic portion comprises a glutamine position 207, position 268, position 317, position 360, position 661, position 694, position 779, and position 876, as numbered according to SEQ ID NO: 6.

[0120] In some embodiments, a Plasmodium P113 polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 136.

[0121] Further provided Is a poiyribonudeotlde encoding a polypeptide, wherein the polypeptide comprises (I) a secretory signal, and (H) a Plasmodium Rlpr polypeptide or antigenic portion thereof.

[0122] In some embodiments, a polypeptide comprises (I) a secretory signal, (II) a Plasmodium Rlpr polypeptide or antigenic portion thereof, and (III) a transmembrane region.

[0123] In some embodiments, a polypeptide comprises (I) a secretory signal, (II) a Plasmodium Rlpr polypeptide or antigenic portion thereof, (ii) a linker, and (hr) a transmembrane region.

[0124] In some embodiments, a secretory signal Is a Plasmodium secretory signal. In some embodiments, a Plasmodium secretory signal Is a Plasmodium Rlpr secretory signal. In some embodiments, a Plasmodium Ripr secretory signal comprises or consists of an amino add sequence according to SEQ ID NO: 186.

[0125] In some embodiments, a secretory signal is a viral seaetory signal. In some embodiments, a secretory signal is an HSV glycoprotein D (gD) secretory signal. In some embodiments, a HSV gD secretory signal comprises or consists of an amino add sequence accorcfing to SEQ ID NO: 42.

[0126] In some embodiments, a Plasmodium Ripr polypeptide or antigenic portion thereof Ripr comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 171. In some embodiments, a Plasmodium Ripr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 171.

[0127] In some embodiments, a Plasmodium Ripr polypeptide or antigenic portion thereof comprises one, two, three, four, five, six, seven, eight, nine, ten, eleven or twelve N-linked glycosylation sites. In some embodiments, a Plasmodium Ripr polypeptide or antigenic portion thereof comprises an amino acid substitution at one or more N- linked glycosylation sites, wherein the amino add substitution prevents glycosylation.

[0128] In some embodiments, a Plasmodium Ripr polypeptide or antigenic portion thereof comprises a substitution of NX[T/S] to QX[T/S]. In some embodiments, a Plasmodium Ripr polypeptide or antigenic portion thereof comprises an amino acid substitution at all of the N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation. [0129] In some embodiments, a Plasmodium Ripr polypeptide or antigenic portion thereof comprises an asparagine at position 103, position 144, position 228, position 303, position 334, position 480, position 498, position 506, position 526, position 646, position 964, position 1021, or a combination thereof, as numbered according to SEQ ID NO: 171. In some embodiments, a Plasmodium Ripr polypeptide or antigenic portion thereof comprises a glutamine at position 103, position 144, position 228, position 303, position 334, position 480, position 498, position 506, position 526, position 646, position 964, position 1021, or a combination thereof, as numbered according to SEQ ID NO: 171. In some embodiments, a Plasmodium Ripr polypeptide or antigenic portion thereof comprises a glutamine at position 103, position 144, position 228, position 303, position 334, position 480, position 498, position 506, position 526, position 646, position 964, and position 1021, as numbered according to SEQ ID NO: 171. In some embodiments, a Plasmodium Ripr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 172.

[0130] In some embodiments, a linker comprises a glycine-serine linker. In some embodiments, a linker comprises or consists of an amino acid sequence according to SEQ ID NO: 138.

[0131] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 145. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 145.

[0132] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 147. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 147.

[0133] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 149. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 149.

[0134] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 151. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 151.

[0135] In some embodiments, a transmembrane region is a heterologous transmembrane region. In some embodiments, a heterologous transmembrane region comprises or consists of a viral transmembrane region. In some embodiments, a heterologous transmembrane region comprises or consists of an HSV transmembrane regon. In some embodiments, a HSV transmembrane region comprises or consists of an HSV gD transmembrane region. In some embodiments, a HSV gD transmembrane region comprises or consists of an amino acid sequence according to SEQ ID NO: 75.

[0136] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 146. In some embodments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 146.

[0137] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 200. In some embodments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 200.

[0138] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 148. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 148. [0139] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 201. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 201.

[0140] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 150. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 150.

[0141] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 202. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 202.

[0142] In some embodiments, a polypeptide comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 152. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 152.

[0143] In some embodiments, a polypeptide comprises or consists of an amino acid sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 203. In some embedments, a polypeptide comprises or consists of an amino acid sequence according to SEQ ID NO: 203.

[0144] In some embodiments, a Plasmodium Ripr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 178.

[0145] In some embodiments, a Plasmodium Ripr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 177. In some embodiments, a Plasmodium Ripr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 177.

[0146] In some embodiments, a Plasmodium Ripr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 176. In some embodiments, a Plasmodium Ripr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 176.

[0147] In some embodiments, a Plasmodium Ripr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 175. In some embodiments, a Plasmodium Ripr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 175.

[0148] In some embodiments, a Plasmodium Ripr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 174. In some embodiments, a Plasmodium Ripr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 174.

[0149] Plasmodium Ripr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 190. In some embodiments, a Plasmodium Ripr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 190.

[0150] In some embodiments, a Plasmodium Ripr polypeptide or antigenic portion thereof comprises an amino acid substitution at one or more N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation. In some embodiments, a Plasmodium Ripr polypeptide or antigenic portion thereof comprises a substitution of NX[T/S] to QX[T/S]. In some embodiments, a Plasmodium Ripr polypeptide or antigenic portion thereof comprises an amino acid substitution at all of the N-linked glycosylation sites, wherein the amino acid substitution prevents glycosylation.

[0151] In some embodiments, a Plasmodium Ripr polypeptide or antigenic portion thereof comprises an asparagine at position 646, position 964, position 1021, or a combination thereof, as numbered according to SEQ ID NO: 190. In some embodiments, a Plasmodium Ripr polypeptide or antigenic portion thereof comprises a glutamine at position 646, position 964, position 1021, or a combination thereof, as numbered according to SEQ ID NO: 190. In some embodiments, a Plasmodium Ripr polypeptide or antigenic portion thereof comprises a glutamine at position 646, position 964, and position 1021, as numbered according to SEQ ID NO: 190. In some embodiments, a Plasmodium Ripr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 173.

[0152] In some embodiments, a polypeptide comprises or consists of an amino acid sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 153. In some embodiments, a polypeptide comprises or consists of an amino acid sequence according to SEQ ID NO: 153.

[0153] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 204. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 204.

[0154] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 154. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 154.

[0155] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 205. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 205.

[0156] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 155. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 155.

[0157] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 206. In some emboefiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 206.

[0158] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 156. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 156.

[0159] In some embodiments, a polypeptide comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 207. In some emboefiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 207.

[0160] In some embodiments, a polypeptide comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 157. In some emboefiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 157. [0161] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 208. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 208.

[0162] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 158. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 158.

[0163] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 209. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 209.

[0164] Further provided Is a polyribonucleotide encoding a polypeptide, wherein the polypeptide comprises (I) a secretory signal, (H) a Plasmodium TRAMP polypeptide or antigenic portion thereof.

[0165] In some embodiments, a secretory signal is a viral secretory signal. In some embodiments, a secretory signal is an HSV glycoprotein D (gD) secretory signal. In some embodiments, a HSV gD secretory signal comprises or consists of an amino add sequence according to SEQ ID NO: 42.

[0166] In some embodiments, a Plasmodium TRAMP polypeptide or antigenic portion thereof comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 179. In some embodiments, a Plasmodium TRAMP polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 179.

[0167] In some embodiments, a Plasmodium TRAMP polypeptide or antigenic portion thereof comprises one, two, three, four, five, six, seven, or eight N-linked glycosylation sites. In some embodiments, a Plasmodium TRAMP polypeptide or antigenic portion thereof comprises an amino acid substitution at one or more N-linked glycosylation sites, wherein the amino acid substitution prevents glycosylation. In some embodiments, a Plasmodium TRAMP polypeptide or antigenic portion thereof comprises a substitution of NX[T/S] to QX[T/S]. In some embodiments, a Plasmodium TRAMP polypeptide or antigenic portion thereof comprises an amino acid substitution at all of the N- linked glycosylation sites, wherein the amino add substitution prevents glycosylation.

[0168] In some embodiments, a Plasmodium TRAMP polypeptide or antigenic portion thereof comprises an asparagine at position 149, position 195, position 202, or a combination thereof, as numbered according to SEQ ID NO: 179. In some embodiments, a Plasmodium TRAMP polypeptide or antigenic portion thereof comprises a glutamine at position 149, position 195, position 202, or a combination thereof, as numbered according to SEQ ID NO: 179. In some embodiments, a Plasmodium TRAMP polypeptide or antigenic portion thereof comprises a glutamine at position 149, position 195, and position 202, as numbered according to SEQ ID NO: 179. In some embodiments, a Pfasmodft/mTRAMP polypeptide or antigenic portion thereof comprises or consists of an amino acid sequence according to SEQ ID NO: 181.

[0169] In some embodiments, a PiasmodiumTRNAP polypeptide or antigenic portion thereof comprises an asparagine at position 112, position 149, position 155, position 170, position 195, position 202, position 253, position 305, or a combination thereof, as numbered according to SEQ ID NO: 179. In some embodiments, a Plasmodium TRAMP polypeptide or antigenic portion thereof comprises a glutamine at position 112, position 149, position 155, position 170, position 195, position 202, position 253, position 305, or a combination thereof, as numbered according to SEQ ID NO: 179. In some embodiments, a Plasmodium TRAMP polypeptide or antigenic portion thereof comprises a glutamine at position 112, position 149, position 155, position 170, position 195, position 202, position 253, and position 305, as numbered according to SEQ ID NO: 179. In some embodiments, a Plasmodium TRAMP polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 180.

[0170] In some embodiments, a Plasmodium TRAMP polypeptide or antigenic portion thereof comprises a PMX cleavage site. In some embodiments, a PMX deavage site comprises or consists of an amino add sequence of HFLQ. In some embodiments, a Plasmodium TRAMP polypeptide or antigenic portion thereof comprises a SUB2 cleavage site. In some embodiments, a SUB2 cleavage site comprises or consists of an amino acid sequence according to SEQ ID NO: 193.

[0171] In some embodiments, a linker comprises a glydne-serine linker. In some embodiments, a linker comprises or consists of an amino acid sequence according to SEQ ID NO: 138.

[0172] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 159. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 159.

[0173] In some embodiments, a polypeptide comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 160. In some embodiments, a polypeptide comprises or consists of an amino acid sequence according to SEQ ID NO: 160.

[0174] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 161. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 161.

[0175] Futher provided is a polyribonudeotkie encoding a polypeptide, wherein the polypeptide comprises (I) a secretory signal, and (li) a Plasmodium CSS polypeptide or antigenic portion thereof.

[0176] In some embodiments, a polypeptide comprises (I) a secretory signal, (ii) a Plasmodium CSS polypeptide or antigenic portion thereof, and (ill) a transmembrane region.

[0177] In some embodiments, a polypeptide comprises (I) a secretory signal, (il) a Plasmodium CSS polypeptide or antigenic portion thereof, (Hi) a linker, and (iv) a transmembrane region.

[0178] In some embodiments, a secretory signal is a Plasmodium secretory signal. In some embodiments, a Plasmodium secretory signal Is a Plasmodium CSS secretory signal. In some embodiments, a Plasmodium CSS secretory signal comprises or consists of an amino add sequence according to SEQ ID NO: 188.

[0179] In some embodiments, a secretory signal is a viral secretory signal. In some embodiments, a secretory signal is an HSV glycoprotein D (gD) secretory signal. In some embodiments, a HSV gD secretory signal comprises or consists of an amino add sequence according to SEQ ID NO: 42.

[0180] In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 182. In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 182.

[0181] In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises one, two, three, four, five, or six N-linked glycosylation sites. In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises an amino add substitution at one or more N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation. In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises a substitution of NX[T/S] to QX[T/S]. In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises an amino add substitution at all of the N-linked glycosylation sites, wherein the amino acid substitution prevents glycosylation.

[0182] In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises an asparagine at position 74, position 88, position 192, position 234, position 261, position 283, or a combination thereof, as numbered according to SEQ ID NO: 182. In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises a glutamine at position 74, position 88, position 192, position 234, position 261, position 283, or a combination thereof, as numbered according to SEQ ID NO: 182. In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises a glutamine at position 74, position 88, position 192, position 234, position 261, and position 283, as numbered according to SEQ ID NO: 182. In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 183.

[0183] In some embodiments, a linker comprises a glydne-serine linker. In some embodiments, a linker comprises or consists of an amino acid sequence according to SEQ ID NO: 138.

[0184] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 162. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 162.

[0185] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 163. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 163.

[0186] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 164. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 164.

[0187] In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 194. In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 194.

[0188] In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 195. In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 195.

[0189] In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 182. In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 182.

[0190] In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises one, two, three, four, five, or six N-linked glycosylation sites. In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises an amino add substitution at one or more N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation. In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises a substitution of NX[T/S] to QX[T/S]. In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises an amino acid substitution at all of the N-linked glycosylation sites, wherein the amino acid substitution prevents glycosylation.

[0191] In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises an asparagine at position 74, position 88, or a combination thereof, as numbered according to SEQ ID NO: 194. In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises a glutamine at position 74, position 88, or a combination thereof, as numbered according to SEQ ID NO: 194. In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises a glutamine at position 74 and position 88, as numbered according to SEQ ID NO: 194. In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises or consists of an amino acid sequence according to SEQ ID NO: 184.

[0192] In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises an asparagine at position 192, position 234, position 261, position 283, or a combination thereof, as numbered according to SEQ ID NO: 195. In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises a glutamine at position 192, position 234, position 261, position 283, or a combination thereof, as numbered according to SEQ ID NO: 195. In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises a glutamine at position 192, position 234, position 261, and position 283, as numbered according to SEQ ID NO: 195. In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 185.

[0193] In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises an asparagine at position 74, position 88, position 192, position 234, position 261, position 283, or a combination thereof, as numbered according to SEQ ID NO: 182. In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises a glutamine at position 74, position 88, position 192, position 234, position 261, position 283, or a combination thereof, as numbered according to SEQ ID NO: 182. In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises a glutamine at position 74, position 88, position 192, position 234, position 261, and position 283, as numbered according to SEQ ID NO: 182. In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises or consists of an amino acid sequence according to SEQ ID NO: 183.

[0194] In some embodiments, a first linker and/or second linker comprises or consists of an amino add sequence according to SEQ ID NO: 138.

[0195] In some embodiments, a transmembrane region is a heterologous transmembrane region. In some embodiments, a heterologous transmembrane region comprises or consists of a viral transmembrane region. In some embodiments, a heterologous transmembrane region comprises or consists of an HSV transmembrane region. In some embodiments, a HSV transmembrane region comprises or consists of an HSV gD transmembrane region. In some embodiments, a HSV gD transmembrane region comprises or consists of an amino add sequence accorcSng to SEQ ID NO: 75.

[0196] In some embodiments, a linker comprises a glydne-serine linker. In some embodiments, a linker comprises or consists of an amino acid sequence according to SEQ ID NO: 138.

[0197] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 165. In some embodiments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 165. [0198] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 166. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 166.

[0199] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 167. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 167.

[0200] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 168. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 168.

[0201] In some embodiments, a polypeptide comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 210. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 210.

[0202] In some embodiments, a polypeptide comprises or consists of an amino acid sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 211. In some embedments, a polypeptide comprises or consists of an amino acid sequence according to SEQ ID NO: 211.

[0203] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 212. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 212.

[0204] In some embodiments, a polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 213. In some embedments, a polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 213.

[0205] In some embodiments, an RNA construct comprises, In 5' to 3' order (I) a 5' UTR, (li) a polyribonucleotide described herein, (ill) a 3' UTR, and (Iv) a polyA tall sequence.

[0206] In some embodiments, an RNA construct comprises (i) a 5' UTR that comprises or consists of a modified human alpha-globin 5 -UTR; and (ii) a 3' UTR that comprises or consists of a first sequence from the amino terminal enhancer of split (AES) messenger.

[0207] In some embodiments, an RNA construct comprises a 5' UTR. In some embodiments, a 5' UTR comprises or consists of a modified human alpha-globin 5 -UTR. In some embodiments, a 5' UTR consists of a ribonudeic add sequence according to SEQ ID NO: 111.

[0208] In some embodiments, an RNA construct comprises a 3' UTR. In some embodiments, a 3' UTR that comprises or consists of a first sequence from the amino terminal enhancer of split (AES) messenger RNA and a second sequence from the mitochondrial encoded 12S ribosomal RNA. In some embodiments, a 3’ UTR consists of a ribonudeic add sequence according to SEQ ID NO: 117.

[0209] In some embodiments, an RNA construct comprises a polyA tail sequence. In some embodiments, a polyA tail sequence is a split polyA tail sequence. In some embodiments, a split polyA tail sequence consists of a ribonudeic add sequence according to SEQ ID NO: 114.

[0210] In some embodiments, an RNA construct comprises a 5* cap. [0211] In some embodiments, an RNA construct comprises a cap proximal sequence comprising positions +1,

+2, +3, +4, and +5 of the polyribonudeotide. [0212] In some embodiments, a 5’ cap comprises or consists of m7(3'OMeG)(5’)ppp(5')(2^,OMeAi)pGz, wherein Ai is position +1 of the polyribonucleotide, and Gz is position +2 of the polyribonucleotide.

[0213] In some embodiments, a cap proximal sequence comprises Ai and Gz of the Capl structure, and a sequence comprising: A3A4U5 (SEQ ID NO: 138) at positions +3, +4 and +5 respectively of the polyribonucleotide. [0214] In some embodiments, a polyribonucleotide provided herein indudes modified uridines in place of all uridines. In some embodiments, modified uridines are each Nl-methyl-pseudouridine.

[0215] The present disdosure also provides compositions. In some embodiments, a composition comprises one or more polyribonucleotides provided herein. In some embodiments, a composition comprises one or more RNA constructs provided herein.

[0216] In some embodiments, a composition comprises (i) one or more polyribonucleotides and (II) lipid nanopartides, polyptexes (PLX), lipidated polyptexes (LPLX), or liposomes. In some embodiments, the one or more polyribonudeotldes are fully or partially encapsulated within the lipid nanopartides, polyplexes (PLX), lipidated polyplexes (LPLX), or liposomes.

[0217] In some embodiments, a composition comprises (i) one or more polyribonudeotldes and (li) lipid nanopartides. In some embodiments, the one or more polyribonucleotides are fully or partially encapsulated within the lipid nanoparticles.

[0218] The present disdosure provides pharmaceutical compositions. In some embodiments, a pharmaceutical composition comprises (I) one or more polyribonucleotides as described herein, one or more RNA constructs as described herein, or a composition as described herein, and (II) at least one pharmaceutically acceptable exdpient [0219] In some embodiments, a pharmaceutical comprises a cryoprotectant. In some embodiments, a cryoprotectant Is sucrose.

[0220] In some embodiments, a pharmaceutical comprises an aqueous buffered solution, optionally wherein the aqueous buffered solution comprises one or more of Tris base, Tris HCI, NaCI, KCI, NazHPCU, and KHzPO«.

[0221] The present disdosure provides, among other things, a combination comprising (i) a first pharmaceutical composition comprising a first polyribonudeotide, wherein the first polyribonucleotide encodes a first polypeptide, and the first polypeptide comprises one or more Plasmodium Rh5 polypeptides or antigenic portions thereof, and (ii) a second pharmaceutical composition comprising a second polyribonudeotide, wherein the second polyribonucleotide encodes a second polypeptide, the second polypeptide comprises one or more Plasmodium Rh5 invasion complex polypeptides selected from one or more CyRPA polypeptides or antigenic portions thereof, one or more Ripr polypeptides or antigenic portions thereof, one or more P113 polypeptides or antigenic portions thereof, one or more TRAMP polypeptides or antigenic portions thereof, one or more CSS polypeptides or antigenic portions thereof, or a combination thereof.

[0222] In some embodiments, a second polypeptide comprises one or more one or more CyRPA polypeptides or antigenic portions thereof. In some embodiments, a second polypeptide comprises one or more one or more Pl 13 polypeptides or antigenic portions thereof. In some embodiments, a second polypeptide comprises one or more one or more Ripr polypeptides or antigenic portions thereof. In some embodiments, a second polypeptide comprises one or more one or more TRAMP polypeptides or antigenic portions thereof. In some embodiments, a second polypeptide comprises one or more one or more CSS polypeptides or antigenic portions thereof.

[0223] The present disclosure also provides a combination comprising (i) a first pharmaceutical composition comprising a first polyribonudeotide, wherein the first polyribonudeotide encodes a first polypeptide, and the first pdypeptide comprises one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof; and (ii) a second pharmaceutical composition comprising a second polyribonucleotide, wherein the second polyribonucleotide encodes a second polypeptide, the second polypeptide comprises one or more PiasmodiumT cell antigens.

[0224] Further, the present disclosure provides a combination comprising: (i) a first pharmaceutical composition comprising a first polyribonucleotide encoding a first polypeptide, and the first polypeptide comprises one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof; and (ii) a second pharmaceutical composition comprising a second polyribonucleotide, wherein the second polyribonucleotide encodes a second polypeptide, the second polypeptide comprises one or more Plasmodium CSV polypeptides or antigenic portions thereof.

[0225] Among other things, a method comprising administering a polyribonucleotide as described herein to a subject. The present disclosure also provides a method comprising administering an RNA construct described herein to a subject. The present disclosure further provides a method comprising administering a composition described herein to a subject. Additionally, a method comprising administering one or more doses of the pharmaceutical composition described herein to a subject.

[0226] In some embodiments, a pharmaceutical composition as provided herein Is for use in the treatment of a malaria Infection comprising administering one or more doses of the pharmaceutical composition to a subject.

[0227] In some embodiments, a pharmaceutical composition as provided herein Is for use in the prevention of a malaria Infection comprising administering one or more doses of the pharmaceutical composition to a subject [0228] In some embodiments, administering two or more doses of the pharmaceutical conrposltlon to a subject. In some embodiments, administering three or more doses of the pharmaceutical composition to a subject

[0229] In some embodiments, a method provided herein comprises administering a combination as described herein to a subject.

[0230] In some embodiments, a first pharmaceutical composition and the second pharmaceutical composition are administered on the same day. In some embodiments, a first pharmaceutical composition and the second pharmaceutical composition are administered on different days. In some embodiments, a pharmaceutical composition and the second pharmaceutical composition are administered to the subject at (Afferent locations on the subjects body.

[0231] In some embodiments, a method is a method of treating a malaria infection.

[0232] In some embodiments, a method is a method of preventing a malaria infection.

[0233] In some embodiments, a subject has or is at risk of developing a malaria infection.

[0234] In some embodiments, a subject is a human.

[0235] In some embodiments, administration induces an anti-malaria immune response in the subject. In some embodiments, an anti-malaria immune response in the subject comprises an adaptive immune response. In some embodiments, an anti-malaria immune response in the subject comprises a T-cell response. In some embodiments, a T-cell response is or comprises a CD4+ T cell response. In some embodiments, a T-cell response is or comprises a CD6+ T cell response. In some embodiments, an anti-malaria immune system response comprises a B-cell response. In some embodiments, an anti-malaria immune system response comprises the production of antibodies directed against the one or more Plasmodium antigens. [0236] The present disdosure provides a use of the pharmaceutical composition as described herein in the treatment of a malaria infection.

[0237] The present disdosure provides a use of the pharmaceutical composition as described herein in the prevention of a malaria infection.

[0238] The present disdosure provides a use of the pharmaceutical composition as described herein in inducing an anti-malaria immune response in a subject.

[0239] Additionally, the present disdosure provides a polypeptide encoded by a polyribonucleotide described herein or an RNA construct described herein.

[0240] The present disclosure also provides a host cell comprising a polyribonucleotide described herein or an RNA construct described herein. The present disclosure further provides a host cell comprises a polypeptide described herein.

BRIEF DESCRIPTION OF THE DRAWING

[0241] FIG. 1 provides an annotated polypeptide corresponding to the P. falciparum Rh5 sequence set out in SEQ ID NO: 1. Certain sequence features are annotated, including secretory signal sequence (red bold corresponds to the signal peptide as described by Baum et al. 2009, the additional 2 red residues are included in the signal peptide description by Wright et al. 2014), glycosylation sites (NXS/T, blue and bold, lower prediction scores in blue only, the motif in brackets is proximal to the basigin binding site), Rh5 PMX cleavage site (black bold), the 3D7 reference sequence has a cysteine at position 203 (red) which is often a tyrosine in naturaNy circulating parasite isolates, paired cysteines are color coded accordingly, and underlined regions are ordered and form the alpha helical composition of the kite structure of the protein, which is associated with basigin binding.

[0242] FIG. 2, parts (A) through (M), includes schematics of exemplary polypeptides encoded by poiyribonudeotkles provided herein. All amino add (aa) references induded refer to the P. falciparum Rh5 sequence set out in SEQ ID NO: 1. (A) includes a schematic of an exemplary polypeptide, RNA Construct 1, encoded by a polyribonudeotlde described herein. RNA Construct 1 comprises an KSV glycoprotein D (gD) secretory signal (e.g., SEQ ID NO: 42), an Rh5 N-terminal "N" disordered region (aa 25-139), an Rh5 ordered region (aa 140-247), an Rh5 linking "L" disordered region (aa 248-296), and an Rh5 ordered region (aa 297-526). Four glycosylation sites at positions 38, 284, 214, and 297 have been mutated from N-X-[T/S] to N-X-A (where X is not proline). (B) indudes a schematic of an exemplary polypeptide, RNA Construct 2, encoded by a polyribonucleotide described herein. RNA Construct 2 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), an Rh5 ordered region (aa 140-247) comprising a C203Y mutation, an Rh5 ordered region (aa 297-526), a glydne-serine linker (e.g., SEQ ID NO: 86), and a foldon domain. (C) indudes a schematic of an exemplary polypeptide, RNA Construct 3, encoded by a polyribonudeotlde described herein. RNA Construct 3 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), an Rh5 ordered region (aa 140-247) comprising a C203Y mutation, and an Rh5 ordered region (297-526). (D) indudes a schematic of an exemplary polypeptide, RNA Construct 4, encoded by a polyribonucleotide described herein. RNA Construct 4 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), an Rh5 ordered region (aa 140-247) comprising a C203Y mutation, an Rh5 ordered region (297-526), a glydne-serine linker (e.g., SEQ ID NO: 137), and a ferritin domain. (E) Indudes a schematic of an exemplary polypeptide, RNA Construct 5, encoded by a polyribonudeotlde described herein. Membrane RNA Construct 5 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), an Rh5 ordered region (aa 140-247) comprising a C203Y mutation, an Rh5 ordered region (297-526), a glydne-serine linker (e.g., SEQ ID NO: 86), and an HSV gD transmembrane region (e.g., SEQ ID NO: 75). (F) indudes a schematic of an exemplary polypeptide, RNA Construct 6, encoded by a polyribonucleotide described herein. RNA Construct 6 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), an Rh5 ordered region (aa 140-247) comprising a C203Y mutation and an N214Q mutation, an Rh5 ordered region (aa 297-526) comprising an N297Q mutation, a glycine-serine linker (e.g., SEQ ID NO: 86), and a foldon domain. (G) indudes a schematic of an exemplary polypeptide, RNA Construct 7, encoded by a polyribonucleotide described herein. RNA Construct 7 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), an Rh5 ordered region (aa 140-247) comprising a C203Y mutation and an N214Q mutation, an Rh5 ordered region (aa 297-526) comprising an N297Q mutation. (H) indudes a schematic of an exemplary polypeptide, RNA Construct 8, encoded by a polyribonucleotide described herein. Self assembling RNA Construct 8 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), an Rh5 ordered region (aa 140-247) comprising a C203Y mutation and an N214Q mutation, an Rh5 ordered region (aa 297- 526) comprising an N297Q mutation, a glydne-serine linker (e.g., SEQ ID NO: 137), and a ferritin domain. (I) Indudes a schematic of an exemplary polypeptide, RNA Construct 9, encoded by a polyribonucleotide described herein. RNA Construct 9 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), an Rh5 ordered region (aa 140-247) comprising a C203Y mutation and an N214Q mutation, an Rh5 ordered region (aa 297-526) comprising an N297Q mutation, a glydne-serine linker (e.g., SEQ ID NO: 86), and an HSV gD transmembrane region (e.g., SEQ ID NO: 75). (J) indudes a schematic of an exemplary polypeptide, RNA Construct 10, encoded by a polyribonucleotide described herein. RNA Construct 10 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), an Rh5 ordered region (aa 140-247) comprising a C203Y mutation and an N214Q mutation, an Rh5 ordered region (aa 297-526), a glydne-serine linker (e.g., SEQ ID NO: 86), and a foldon domain. (K) indudes a schematic of an exemplary polypeptide, RNA Construct 11, encoded by a polyribonucleotide described herein. RNA Construct 11 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), an Rh5 ordered region (aa 140-247) comprising a C203Y mutation and an N214Q mutation, an Rh5 ordered region (aa 297-526). (L) Indudes a schematic of an exemplary polypeptide, RNA Construct 12, encoded by a polyribonucleotide described herein. RNA Construct 12 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), an Rh5 ordered region (aa 140-247) comprising a C203Y mutation and an N214Q mutation, an Rh5 ordered region (aa 297-526), a glycine-serine linker (e.g., SEQ ID NO: 137), and a ferritin domain. (M) includes a schematic of an exemplary polypeptide, RNA Construct 13, encoded by a polyribonudeotkie described herein. RNA Construct 13 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), an Rh5 ordered region (aa 140-247) comprising a C203Y mutation and an N214Q mutation, an Rh5 ordered region (aa 297-526), a glydne-serine linker (e.g., SEQ ID NO: 86), and an HSV gD transmembrane region (e.g., SEQ ID NO: 75).

[0243] FIG. 3 provides an annotated polypeptide corresponding to the P. falciparum CyRPA sequence set out In SEQ ID NO: 3. Certain sequence features are annotated, Indudlng secretory signal sequence (red) and glycosylation sites (NXS/T, blue).

[0244] FIG. 4, parts (A) through (F), includes schematics of exemplary polypeptide encoded by polyribonudeotides provided herein. All amino add (aa) references induded refer to the P. falciparum C/RPk sequence set out in SEQ ID NO: 3. (A) includes a schematic of an exemplary polypeptide, RNA Construct 14, encoded by a polyribonudeotkie described herein. RNA Construct 14 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of CyRPA (aa 30-362), a glydne-serine linker (e.g., SEQ ID NO: 86), and a foldon tag. (B) indudes a schematic of an exemplary polypeptide, RNA Construct 15, encoded by a polyribonucleotide described herein. RNA Construct IScomprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), and a portion of CyRPA (aa 30-362). (C) includes a schematic of an exemplary polypeptide, RNA Construct 16, encoded by a polyribonucleotide described herein. RNA Construct 16 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of CyRPA (aa 30-362), a glydne-serine linker (e.g., SEQ ID NO: 86), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75). (D) includes a schematic of an exemplary polypeptide, RNA Construct 17, encoded by a polyribonucleotide described herein. RNA Construct 17 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of CyRPA (aa 30-362) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at three glycosylation sites (e.g., at amino acid positions 145, 332, and 338, as numbered according to SEQ ID NO: 3), a glycine-serine linker (e.g., SEQ ID NO: 86), and a foidon tag. (E) includes a schematic of an exemplary polypeptide, RNA Construct 18, encoded by a polyribonucleotide described herein. RNA Construct 18 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), and a portion of CyRPA (aa 30-362) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X Is not proline) at three glycosylation sites. (F) includes a schematic of an exemplary polypeptide, RNA Construct 19, encoded by a polyribonudeotlde described herein. RNA Construct 19 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of CyRPA (aa 30-362) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at three glycosylation sites, a glycine-serine linker (e.g., SEQ ID NO: 86), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).

[0245] FIG. 5, parts (A) through (G), Includes schematics of exemplary polypeptide encoded by polyribonucleotides provided herein. All amino add (aa) references Included refer to the P. fatoparum CyRPA sequence set out h SEQ ID NO: 3. Polypeptides may optionally Include one or more linkers between various regions and/or domains. (A) Indudes a schematic of an exemplary polypeptide, RNA Construct 22, encoded by a polyribonudeotlde described herein. RNA Construct 22 comprises CyRPA secretory signal (SEQ ID NO: 187) and a portion of CyRPA (aa 29-362). (B) indudes a schematic of an exemplary polypeptide, RNA Construct 23, encoded by a polyribonucleotide described herein. RNA Construct 23 comprises a CyRPA secretory signal (SEQ 10 NO: 187) and a portion of CyRPA (aa 29-362) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at three glycosylation sites (e.g., at amino add positions 145, 332, and 338, as numbered according to SEQ ID NO: 3). (C) indudes a schematic of an exemplary polypeptide, RNA Construct 24, encoded by a polyribonudeotlde described herein. RNA Construct 24 comprises an HSV gD secretory signal (e.g., SEQ ID NO:42) and a portion of CyRPA (aa 29- 362) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at three glycosylation sites (e.g., at amino add positions 145, 332, and 338, as numbered according to SEQ ID NO: 3). (D) includes a schematic of an exemplary polypeptide, RNA Construct 25, encoded by a polyribonucleotide described herein. RNA Construct 25 comprises a full-length CyRPA (aa 1-362) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at three glycosylation sites (e.g., at amino add positions 145, 332, and 338, as numbered according to SEQ ID NO: 3) and a HSV gD transmembrane region (e.g., SEQ ID NO: 75). (E) Indudes a schematic of an exemplary polypeptide, RNA Construct 26, encoded by a polyribonucleotide described herein. RNA Construct 26 comprises an HSV gD secretory signal (e.g., SEQ ID NO:42), a portion of CyRPA (aa 29-362) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at three glycosylation sites (e.g., at amino add positions 145, 332, and 338, as numbered according to SEQ ID NO: 3) and a HSV gD transmembrane region (e.g., SEQ ID NO: 75). (F) includes a schematic of an exemplary polypeptide, RNA Construct 51, encoded by a polyribonucleotide described herein. RNA Construct 51 comprises a full-length CyRPA (aa 1-362) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at three glycosylation sites (e.g., at amino add positions 145, 332, and 338, as numbered according to SEQ ID NO: 3), a glycine-serine linker (e.g., SEQ ID NO: 138), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75). (G) indudes a schematic of an exemplary polypeptide, RNA Construct 52, encoded by a polyribonudeotlde described herein. RNA Construct 52 comprises an HSV gD secretory signal (e.g., SEQ ID NO:42), a portion of CyRPA (aa 29- 362) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at three glycosylation sites (e.g., at amino add positions 145, 332, and 338, as numbered according to SEQ ID NO: 3), a glycine-serine linker (e.g., SEQ ID NO: 138), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).

[0246] FIG. 6, parts (A) and (8) includes schematics of exemplary polypeptide encoded by pdyribonudeotides provided herein. All amino add (aa) references included refer to the P. falciparum P113 sequence set out in SEQ ID NO: 6. (A) includes a schematic of an exemplary polypeptide, RNA Construct 20, encoded by a polyribonudeotlde described herein. RNA Construct 20 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of P113 (aa 23-939), a glydne-serine linker (e.g., SEQ ID NO: 86), and an HSV gD transmembrane region (e.g., SEQ ID NO: 75). (B) includes a schematic of an exemplary polypeptide, RNA Construct 21, encoded by a polyribonudeotlde described herein. RNA Construct 21 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of P113 (aa 23-939) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at eight glycosylation sites (e.g., at amino add positions 207, 268, 317, 360, 661, 697, 779, 876, 938, as numbered according to SEQ ID NO: 6), a glycine-serine linker (e.g., SEQ ID NO: 86), and an HSV gD transmembrane region (e.g., SEQ ID NO: 75).

[0247] FIG. 7 provides an annotated polypeptide corresponding to the A falciparum Ripr sequence set out In SEQ ID NO: 2. Certain sequence features are annotated, including secretory signal sequence (red), glycosylation sites (NXS/T, blue, stronger predictions in bold), Ripr PMX cleavage site (green), epidermal growth factor domains as described by Chen et al. 2011 are underlined, and the domain In bold Is EGF 7 which has been associated with driving the strongest growth Inhibitory responses (Healer 2019, Nagaoka 2020).

[0248] FIG. 8, parts (A) through (L) Includes schematics of exemplary polypeptides encoded by pdyribonudeotides provided herein. All amino add (aa) references included refer to the P. falciparum Ripr sequences set out in SEQ ID NO: 2. Polypeptides may optionally Include one or more linkers between various regions and/or domains. (A) includes a schematic of an exemplary polypeptide, RNA Construct 27, encoded by a polyribonudeotlde described herein. RNA Construct 27 comprises a Ripr secretory signal (SEQ ID NO: 186) and a portion of Ripr (aa 20- 1086), a glycine-serine linker (e.g., SEQ ID NO: 138). (B) includes a schematic of an exemplary polypeptide, RNA Construct 28, encoded by a polyribonudeotlde described herein. RNA Construct 28 comprises a Ripr secretory signal (SEQ ID NO: 186), a portion of Ripr (aa 20-1086) and a HSV gD transmembrane region (e.g., SEQ ID NO: 75). (C) indudes a schematic of an exemplary polypeptide, RNA Construct 29, encoded by a polyribonudeotlde described herein. RNA Construct 29 comprises a Ripr secretory signal (SEQ ID NO: 186) and a portion of Ripr (aa 20-1086) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X Is not proline) at twelve glycosylation sites (e.g., at amino add positions 103, 144, 228, 303, 334, 480, 498, 506, 526, 646, 964, and 1021 as numbered according to SEQ ID NO: 2). (D) indudes a schematic of an exemplary polypeptide, RNA Construct 30, encoded by a polyribonucleotide described herein. RNA Construct 30 comprises a Ripr secretory signal (SEQ ID NO: 186), a portion of Ripr (aa 20- 1086) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at twelve glycosylation sites (e.g., at amino acid positions 103, 144, 228, 303, 334, 480, 498, 506, 526, 646, 964, and 1021 as numbered according to SEQ ID NO: 2), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75). (E) indudes a schematic of an exemplary polypeptide, RNA Construct 31, encoded by a polyribonucleotide described herein. RNA Construct 31 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42) and a portion of Ripr (aa 20-1086). (F) indudes a schematic of an exemplary polypeptide, RNA Construct 32, encoded by a polyribonucleotide described herein. RNA Construct 32 comprises a an HSV gD secretory signal (e.g., SEQ ID NO:42), a portion of Ripr (aa 20-1086), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75). (G) includes a schematic of an exemplary polypeptide, RNA Construct 33, encoded by a polyribonucleotide described herein. RNA Construct 33 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42) and a portion of Ripr (aa 20-1086) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at twelve glycosylation sites (e.g., at amino acid positions 103, 144, 228, 303, 334, 480, 498, 506, 526, 646, 964, and 1021 as numbered according to SEQ ID NO: 2). (H) includes a schematic of an exemplary polypeptide, RNA Construct 34, encoded by a polyribonucleotide described herein. RNA Construct 34 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of Ripr (aa 20-1086) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at twelve glycosylation sites (e.g., at amino add positions 103, 144, 228, 303, 334, 480, 498, 506, 526, 646, 964, and 1021 as numbered according to SEQ ID NO: 2), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75). (I) hdudes a schematic of an exemplary polypeptide, RNA Construct

53, encoded by a polyribonucleotide described herein. RNA Construct 53 comprises a Ripr secretory signal (SEQ ID NO: 186), a portion of Ripr (aa 20-1086), a glydne-serine linker (e.g., SEQ ID NO: 138), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75). (J) indudes a schematic of an exemplary polypeptide, RNA Construct

54, encoded by a polyribonucleotide described herein. RNA Construct 54 comprises a Ripr secretory signal (SEQ ID NO: 186), a portion of Ripr (aa 20-1086) comprising N-X-(T/S] to Q-X-[T/S] mutations (where X Is not proline) at twelve glycosylation sites (e.g., at amino acid positions 103, 144, 228, 303, 334, 480, 498, 506, 526, 646, 964, and 1021 as numbered according to SEQ JD NO: 2), a glydne-serine linker (e.g., SEQ ID NO: 138), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75). (K) Includes a schematic of an exemplary polypeptide, RNA Construct

55, encoded by a polyribonudeotide described herein. RNA Construct 55 comprises a an HSV gD secretory signal (e.g., SEQ ID NO:42), a portion of Ripr (aa 20-1086), a glydne-serine linker (e.g., SEQ ID NO: 138), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75). (L) Includes a schematic of an exemplary polypeptide, RNA Construct

56, encoded by a polyribonudeotide described herein. RNA Construct 56 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of Ripr (aa 20-1086) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at twelve glycosylation sites (e.g., at amino add positions 103, 144, 228, 303, 334, 480, 498, 506, 526, 646, 964, and 1021 as numbered according to SEQ ID NO: 2), a glydne-serine linker (e.g., SEQ ID NO: 138), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).

[0249] FIG. 9, parts (A) through (L) includes schematics of exemplary polypeptides encoded by pdyribonudeotides provided herein. All amino add (aa) references included refer to the P. falciparum Ripr sequences set out in SEQ ID NO: 2. Polypeptides may optionally indude one or more linkers between various regions and/or domains. (A) includes a schematic of an exemplary polypeptide, RNA Construct 35, encoded by a polyribonudeotide described herein. RNA Construct 35 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of Ripr (aa 560-1086) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at twelve glycosylation sites (e.g., at amino add positions 646, 964, and 1021 as numbered according to SEQ ID NO: 2), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75). (B) indudes a schematic of an exemplary polypeptide, RNA Construct 36, encoded by a polyribonudeotide described herein. RNA Construct 36 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of Ripr (aa 720-934), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75). (C) indudes a schematic of an exemplary polypeptide, RNA Construct 37, encoded by a polyribonudeotide described herein. RNA Construct 37 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of Ripr (aa 769- 900) and a HSV gD transmembrane region (e.g., SEQ ID NO: 75). (D) indudes a schematic of an exemplary polypeptide, RNA Construct 38, encoded by a polyribonucleotide described herein. Membrane Ripr comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of Ripr (aa 769-856) and a HSV gD transmembrane region (e.g., SEQ ID NO: 75). (E) indudes a schematic of an exemplary polypeptide, Membrane Ripr, encoded by a polyribonudeotide described herein. RNA Construct 38 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of Ripr (aa 817-900), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75). (F) includes a schematic of an exemplary polypeptide, RNA Construct 40, encoded by a polyribonudeotide described herein. RNA Construct 40 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of Ripr (aa 817-856), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75). (G) indudes a schematic of an exemplary polypeptide, RNA Construct 57, encoded by a pdyribonudeotide described herein. RNA Construct 57 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of Ripr (aa 560-1086) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at twelve glycosylation sites (e.g., at amino acid positions 646, 964, and 1021 as numbered according to SEQ ID NO: 2), a glydne-serine linker (e.g., SEQ ID NO: 138), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75). (H) includes a schematic of an exemplary polypeptide, RNA Construct 58, encoded by a pdyribonudeotide described herein. RNA Construct 58 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of Ripr (aa 720-934), a glydne-serine inker (e.g., SEQ ID NO: 138), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75). (I) indudes a schematic of an exemplary polypeptide, RNA Construct 59, encoded by a polyribonucleotide described herein. RNA Construct 59 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of Ripr (aa 769-900), a glydne-serine inker (e.g., SEQ ID NO: 138), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75). (J) Indudes a schematic of an exemplary polypeptide, RNA Construct 60, encoded by a pdyribonudeotide described herein. RNA Construct 60 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of Ripr (aa 769-856), a glydne-serine inker (e.g., SEQ ID NO: 138), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75). (K) indudes a schematic of an exemplary polypeptide, RNA Construct 61, encoded by a pdyribonudeotide described herein. RNA Construct 61 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of Ripr (aa 817-900), a glydne-serine inker (e.g., SEQ ID NO: 138), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75). (L) Includes a schematic of an exemplary pdypeptlde, RNA Construct 62, encoded by a polyribonucleotide described herein. RNA Construct 62 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a portion of Ripr (aa 817-856), a glydne-serine inker (e.g., SEQ ID NO: 138), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).

[0250] FIG. 10 provides an annotated polypeptide corresponding to the P. fataparum'VV.NAV sequence set out in SEQ ID NO: 4. Certain sequence features are annotated, induding secretory signal sequence (red), glycosylation sites (NXS/T, blue), TRAMP deavage site (bold), and SUB2 deavage site (green).

[0251] FIG. 11, parts (A) through (C) indudes schematics of exemplary polypeptides encoded by pdyribonudeotides provided herein. All amino add (aa) references included refer to the P. faldparum'ttMW sequences set out in SEQ ID NO: 4. Polypeptides may optionally indude one or more linkers between various regions and/or domains. (A) indudes a schematic of an exemplary polypeptide, RNA Construct 41, encoded by a polyribonudeotide described herein. RNA Construct 41 comprises an HSV gD secretory signal (e.g., SEQ ID NO:42) and a portion of TRAMP (aa 42-352). (B) indudes a schematic of an exemplary polypeptide, RNA Construct 42, encoded by a polyribonudeotide described herein. RNA Construct 42 comprises an HSV gD secretory signal (e.g., SEQ ID NO:42) and a portion of TRAMP (aa 42-352) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at eight glycosylation sites (e.g., at amino acid positions, 112, 149, 155, 170, 195, 202, 253, and 305, as numbered according to SEQ ID NO: 4). (C) includes a schematic of an exemplary polypeptide, RNA Construct 43, encoded by a polyribonucleotide described herein. RNA Construct 43 comprises an HSV gD secretory signal (SEQ ID NO:42) and a portion of TRAMP (aa 42-352) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at three glycosylation sites (e.g., at amino acid positions, 149, 195, and 202, as numbered according to SEQ ID NO: 4). [0252] FIG. 12 provides an annotated polypeptide corresponding to the P. falciparum CSS sequence set out in SEQ ID NO: 5. Certain sequence features are annotated, including secretory signal sequence (red), glycosylation sites (NXS/T, blue), domains DI and D2 are underlined and joined by the linker region in purple.

[0253] FIG. 13, parts (A) through (K) includes schematics of exemplary polypeptides encoded by polyribonucleotides provided herein. All amino add (aa) references Included refer to the P. falciparum CSS sequences set out in SEQ ID NO: 5. Polypeptides may optionally indude one or more linkers between various regions and/or domains. (A) includes a schematic of an exemplary polypeptide, RNA Construct 44, encoded by a polyribonudeotide described herein. RNA Construct 44 comprises a CSS secretory signal (SEQ ID NO: 188) and a portion of CSS (aa 21- 290). (B) includes a schematic of an exemplary polypeptide, RNA Construct 45, encoded by a polyribonucleotide described herein. RNA Construct 45 comprises a CSS secretory signal (SEQ ID NO: 188) and a portion of CSS (aa 21- 290) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at six glycosylation sites (e.g., at amino add positions 74, 88, 192, 234, 261, and 283 as numbered according to SEQ ID NO: 5). (C) includes a schematic of an exemplary polypeptide, RNA Construct 46, encoded by a polyribonudeotide described herein. RNA Construct 46 comprises an HSV gD secretory signal (e.g., SEQ ID NO:42) and a portion of CSS (aa 21-290) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at six glycosylation sites (e.g., at amino acid positions 74, 88, 192, 234, 261, and 283 as numbered according to SEQ ID NO: 5). (D) Includes a schematic of an exemplary polypeptide, RNA Construct 47, encoded by a polyribonudeotide described herein. RNA Construct 47 comprises a CSS secretory signal (SEQ ID NO: 188), a portion of CSS (aa 21-290) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at six glycosylation sites (e.g., at amino add positions 74, 88, 192, 234, 261, and 283 as numbered according to SEQ ID NO: 5), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75). (E) indudes a schematic of an exemplary polypeptide, RNA Construct 48, encoded by a polyribonucleotide described herein. RNA Construct 48 comprises an HSV gD secretory signal (e.g., SEQ ID NO:42), a portion of CSS (aa 21-290) comprising N-X-[T/S] to Q- X-[T/S] mutations (where X is not proline) at six glycosylation sites (e.g., at amino add positions 74, 88, 192, 234, 261, and 283 as numbered according to SEQ ID NO: 5), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75). (F) includes a schematic of an exemplary polypeptide, RNA Construct 49, encoded by a polyribonucleotide described herein. RNA Construct 49 comprises an HSV gD secretory signal (e.g., SEQ ID NO:42), a portion of CSS (aa 21-152) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X Is not proline) at two glycosylation sites (e.g., at amino add positions 74 and 88, as numbered according to SEQ ID NO: 5), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75). (G) includes a schematic of an exemplary polypeptide, RNA Construct 50, encoded by a polyribonudeotide described herein. RNA Construct 50 comprises an HSV gD secretory signal (e.g., SEQ ID NO:42), a portion of CSS (aa 153-290) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at four glycosylation sites (e.g., at amino acid positions 192, 234, 261, and 283 as numbered according to SEQ ID NO: 5), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75). (H) indudes a schematic of an exemplary polypeptide, RNA Construct 63, encoded by a polyribonudeotide described herein. RNA Construct 63 comprises a CSS secretory signal (SEQ ID NO: 188), a portion of CSS (aa 21-290) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at six glycosylation sites (e.g., at amino acid positions 74, 88, 192, 234, 261, and 283 as numbered according to SEQ ID NO: 5), a glydne-serine linker (e.g., SEQ ID NO: 138), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75). (I) includes a schematic of an exemplary polypeptide, RNA Construct 64, encoded by a polyribonudeotide described herein. RNA Construct 64 comprises an HSV gD secretory signal (e.g., SEQ ID NO:42), a portion of CSS (aa 21-290) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at six glycosylation sites (e.g., at amino acid positions 74, 88, 192, 234, 261, and 283 as numbered according to SEQ ID NO: 5), a glydne-serine linker (e.g., SEQ ID NO: 138), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75). (J) indudes a schematic of an exemplary polypeptide, RNA Construct 65, encoded by a polyribonucleotide described herein. RNA Construct 65 comprises an HSV gD secretory signal (e.g., SEQ ID NO:42), a portion of CSS (aa 21-152) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X Is not proline) at two glycosylation sites (e.g., at amino add positions 74 and 88, as numbered according to SEQ ID NO: 5), a glydne-serine linker (e.g., SEQ ID NO: 138), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75). (K) indudes a schematic of an exemplary polypeptide, RNA Construct 66, encoded by a polyribonudeotide described herein. RNA Construct 66 comprises an HSV gD secretory signal (e.g., SEQ ID NO:42), a portion of CSS (aa 153-290) comprising N-X-[T/S] to Q-X-[T/S] mutations (where X is not proline) at four glycosylation sites (e.g., at amino add positions 192, 234, 261, and 283 as numbered according to SEQ ID NO: 5), a glydne-serine linker (e.g., SEQ ID NO: 138), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).

[0254] FIG. 14, parts (A) through (F) depicts In-vitro host cell viabllty, transfection rate, and expression of polyribonudeotide constructs encoding Plasmodium polypeptides as described herein by host cells (e.g., HEK293T cells). (A) depicts the percentage of viable host cells that are positive for presence of Intracellularly expressed protein for both transfected and non-transfected (NT) cells. (B) percentage of viable host cells that are positive for presence of surface expressed protein for both transfected and non-transfected cells. (C) depicts transfection rate of Indicated polyribonudeotide constructs as measured by percentage of total host cell population that are positive for presence of Intracellularly expressed protein. (D) depicts transfection rate of Indicated polyribonudeotide constructs as measured by percentage of total host cell population that are positive for presence of surface expressed protein. (E) depicts total Intracellular protein expression as measured by median fluorescence Intensity of the total host cell population for both transfected and non-transfected cells. (F) depicts total surface protein expression as measured by median fluorescence of the total host cell population for both transfected and non-transfected cells. Numbers depicted at x-axis indicate RNA construct number. NT stands for non-transfected.

[0255] FIG. 15, parts (A) through (D) depict transfection rate and expression of polyribonudeotide constructs encoding Plasmodium polypeptides as described herein by host cells (e.g., HEK293T cells). (A) depicts transfection rate of indicated polyribonucleotide constructs as measured by percentage of total host cell population that are positive for presence of intracellularly expressed protein. (B) depicts transfection rate of indicated polyribonucleotide constructs as measured by percentage of total host cell population that are positive for presence of surface expressed protein. (C) depicts total intracellular protein expression as measured by median fluorescence intensity of the total host cell population for both transfected and non-transfected cells. (D) depicts total surface protein expression as measured by median fluorescence of the total host cell population for both transfected and non- transfected cells. Numbers depicted at x-axis indicate RNA construct number. NT stands for non-transfected.

[0256] FIG. 16, parts (A) and (B) depict transfection rate and expression of polyribonucleotide constructs encoding Plasmodium polypeptides as described herein by HEK293T cells. (A) depicts transfection rate of indicated polyribonudeotide constructs as measured by percentage of total host cell popdation that are positive for presence of intracellularly expressed protein. For each construct shown, cells were stained using day 35 serum at 1:1000 dilution from mice immunized twice with the same construct. Cells transfected with construct 23 were stained using serum from mice immunized with construct 23. Construct 1, which encodes RH5, was induded as a control, as ELISA and IVE data with monodonal antibodies have already confirmed immunogenidty for these constructs. Construct 1 was stained with both the monoclonal antibody 9AD4 at 1:2000 dilution and serum from mice immunized with construct 1. (B) depicts total protein expression as measured by median fluorescence intensity of the total host cell population for both non-transfected and transfected cells. For all constructs, un-transfected cells (-) were induded as a negative control, (+) indicates cells that were transfected with the matching construct before staining. Numbers depicted at x-axIs indicate RNA construct number. NT stands for non-transfected.

[0257] FIG. 17 parts (A) through (J) depict in-vitro expression of polyribonudeotlde constructs encoding Plasmodium polypeptides as described herein by host cells (e.g., HEK293T cells). In-vitro expression was measured by detecting the protein associated with cells, either intracellular or surface bound, or the protein detected in the culture medium (e.g., exported). (A) depicts total cell-associated protein expression determined with a HiBit luminescence assay for GARP constructs and (B) depicts total exported protein detected In the culture medium for GARP constructs. (C) depicts total cell-assodated protein expression determined with the HiBit luminescence assay for CyRPA constructs and (D) depicts total exported protein detected in the culture medium for CyRPA constructs. (E) depicts total cell-assodated protein expression determined with the HiBit luminescence assay for Rlpr constructs, and (F) depicts total exported protein detected In the culture medium for Rlpr constructs. (G) depicts total cell- assodated protein expression determined with the HiBit luminescence assay for CSS constructs, and (H) depicts total exported protein detected In the culture medium for CSS constructs. (I) depicts total cell-assodated protein expression determined with the HiBit luminescence assay fbr TRAMP constructs, and (J) depicts total exported protein detected in the culture medium fbr TRAMP constructs. Numbers depicted at x-axis Indicate RNA construct number.

[0258] FIG. 18 depicts ELISA endpoint reciprocal titers for day 35 serum samples of mice Immunized twice with lug of RH5 encoding constructs. Negative control samples are from mice immunized with a construct encoding CSP. Numbers depicted at x-axis indicate RNA construct number.

[0259] FIG. 19, parts (A) through (D), Includes schematics of exemplary polypeptides encoded by polyribonucleotides provided herein. All amino add (aa) references included refer to the P. falciparum GW? sequence set out in SEQ ID NO: 23. (A) indudes a schematic of an exemplary polypeptide, RNA Construct 77, encoded by a polyribonucleotide described herein. RNA Construct 77 comprises an HSV glycoprotein D (gD) secretory signal (e.g., SEQ ID NO: 42) and a GARP-A region (aa 410-673) (SEQ ID NO: 312). (B) indudes a schematic of an exemplary polypeptide, RNA Construct 78, encoded by a polyribonudeotlde described herein. RNA Construct 78 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), and a GARP-A region (aa 410-673) (SEQ ID NO: 312) comprising a N504Q mutation (SEQ ID NO: 313). (C) indudes a schematic of an exemplary polypeptide, RNA Construct 82, encoded by a polyribonucleotide described herein. RNA Construct 82 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a GARP-A region (aa 410-673) (SEQ ID NO: 312) comprising a N504Q mutation (SEQ ID NO: 312), a glydne-serine linker (e.g., SEQ ID NO: 86), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75). (D) indudes a schematic of an exemplary polypeptide, RNA Construct 84, encoded by a polyribonudeotlde described herein. RNA Construct 84 comprises an HSV gD secretory signal (e.g., SEQ ID NO: 42), a GARP-A region (aa 52-673) (SEQ ID NO: 342) comprising N183Q, N233Q, N242Q, and N504Q mutations (SEQ ID NO: 309), and a glycine-serine linker (e.g., SEQ ID NO: 86), and a HSV gD transmembrane region (e.g., SEQ ID NO: 75).

[0260] FIG. 20, parts (A) through (C), includes schematics of exemplary polypeptides encoded by polyribonucleotides provided herein. All amino add (aa) references included refer to the P. faldpanim"VMNP (e.g., PTRAMP) sequence set out in SEQ ID NO: 4. (A) includes a schematic of an exemplary polypeptide, RNA Construct 133, encoded by a polyribonudeotide described herein. RNA Construct 133 comprises an HSV glycoprotein D (gD) secretory signal (e.g., SEQ ID NO: 42) and a TRAMP region (aa 42-352). (B) indudes a schematic of an exemplary polypeptide, RNA Construct 134, encoded by a polyribonudeotide described herein. RNA Construct 134 comprises an HSV glycoprotein D (gD) secretory signal (e.g., SEQ ID NO: 42) and a TRAMP region (aa 42-352) comprising N112Q, N129Q, N155Q, N170Q, N195Q, N202Q, N253Q, and N305Q mutations. (C) Includes a schematic of an exemplary polypeptide, RNA Construct 135, encoded by a polyribonudeotide described herein. RNA Construct 135 comprises an HSV glycoprotein D (gD) secretory signal (e.g., SEQ ID NO: 42) and a TRAMP region (aa 42-352) comprising N149Q, N195Q, and N202Q mutations.

DEFINITIONS

[0261] Compounds of this disclosure include those described generally above and are further illustrated by the classes, subclasses, and species disclosed herein. As used herein, the following definitions shall apply unless otherwise indicated. For purposes of this disclosure, the chemical elements are identified in accordance with the Periodic Table of Elements, CAS version, Handbook of Chemistry and Physics, 75th Ed. Additionally, general principles of organic chemistry are described in “Organic Chemistry", Thomas Sorrell, University Science Books, Sausalito: 1999, and “March's Advanced Organic Chemistry", Sth Ed., Ed.: Smith, M.B. and March, J., John Wiley & Sons, New York: 2001, the entire contents each of which are hereby incorporated by reference.

[0262] Unless otherwise stated, structures depicted herein are meant to include all stereoisomeric (e.g., enantiomeric or diastereomeric) forms of the structure, as well as all geometric or conformational isomeric forms of the structure. For example, the R and S configurations of each stereocenter are contemplated as part of the disclosure. Therefore, single stereochemical Isomers, as well as enantiomeric, diastereomeric, and geometric (or conformational) mixtures of provided compounds are within the scope of the disclosure. For example, in some cases, provided compounds show one or more stereoisomers of a compound, and unless otherwise indicated, represents each stereoisomer alone and/or as a mixture. Unless otherwise stated, all tautomeric forms of provided compounds are within the scope of the disclosure.

[0263] Unless otherwise Indicated, structures depicted herein are meant to Include compounds that differ only In the presence of one or more Isotoplcalty enriched atoms. For example, compounds having the present structures including replacement of hydrogen by deuterium or tritium, or replacement of a carbon by 13C- or 14C-enrkhed carbon are within the scope of this disclosure.

[0264] About The term “about", when used herein in reference to a value, refers to a value that Is similar, in context to the referenced value. In general, those skilled in the art, familiar with the context, will appreciate the relevant degree of variance encompassed by “about” in that context For example, In some embodiments, the term “about” may encompass a range of values that within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less of the referred value. [0265] As used herein, the term "agent,* may refer to a physical entity. In some embodiments, an agent may be characterized by a particular feature and/or effect. For example, as used herein, the term "therapeutic agent' refers to a physical entity has a therapeutic effect and/or elicits a desired biological and/or pharmacological effect In some embodiments, an agent may be a compound, molecule, or entity of any chemical class including, for example, a small molecule, polypeptide, nucleic add, saccharide, lipid, metal, or a combination or complex thereof. [0266] Amino add: In its broadest sense, as used herein, the term "amino acid” refers to a compound and/or substance that can be, is, or has been incorporated into a polypeptide chain, e.g., through formation of one or more peptide bonds. In some embodiments, an amino acid has the general structure H2N-C(H)(R)-C00H. In some embodiments, an amino add is a naturally-occurring amino acid. In some embodiments, an amino acid is a nonnatural amino acid; in some embodiments, an amino add is a D-amlno add; In some embodiments, an amino acid Is an L-amlno add. "Standard amino acid* refers to any of the twenty standard L-amino acids commonly found in naturally occurring peptides. "Nonstandard amino add* refers to any amino add, other than the standard amino acids, regardless of whether it is prepared synthetical or obtained from a natural source. In some embodiments, an amino acid, including a carboxy- and/or amino-terminal amino add in a polypeptide, can contain a structural modification as compared with the general structure above. For example, in some embodiments, an amino add may be modified by methylation, amidation, acetylation, pegylation, glycosylation, phosphorylation, and/or substitution (e.g., of the amino group, the carboxylic add group, one or more protons, and/or the hydroxyl group) as compared with the general structure. In some embodiments, such modification may, for example, alter the circidating half-life of a polypeptide containing the modified amino acid as compared with one containing an otherwise identical unmodified amino add. In some embodiments, such modification does not significantly alter a relevant activity of a polypeptide containing the modified amino add, as compared with one containing an otherwise Identical unmodified amino add. As will be dear from context, in some embodiments, the term "amino acid' may be used to refer to a free amino add; in some embodiments it may be used to refer to an amino add residue of a polypeptide.

[0267] Antigen.-The term "antigen”, as used herein, refers to an agent that (I) elicits an Immune response; and/or (ii) an agent that binds to a T cell receptor (e.g_v when presented by an MHC molecule) or to an antibody. [0268] Anti-malaria immuno response. The term "anti-malaria immune response", as used herein, refers to an Immune response directed to one or more antigens derived from Plasmodium.

[0269] Assodated. Two events or entities are "associated" with one another, as that term is used herein, if the presence, level, degree, type and/or form of one is correlated with that of the other. For example, a particular entity (e.g., polypeptide, genetic signature, metabolite, microbe, etc.) is considered to be assodated with a particular disease, disorder, or condition, if its presence, level and/or form correlates with incidence of, susceptibility to, severity of, stage of, etc. the disease, disorder, or condition (e.g., across a relevant population). In some embodiments, two or more entities are physically "assodated” with one another if they interact, directly or indirectly, so that they are and/or remain in physical proximity with one another. In some embodiments, two or more entities that are physically associated with one another are covalently linked to one another; in some embodiments, two or more entities that are physically associated with one another are not covalently linked to one another but are non- covalently associated, for example by means of hydrogen bonds, van der Waals interaction, hydrophobic interactions, magnetism, and combinations thereof.

[0270] Characteristic portion. As used herein, the term "characteristic portion', In the broadest sense, refers to a portion of a polypeptide or region thereof whose presence (or absence) correlates with presence (or absence) of a particular feature, attribute, or activity of the polypeptide or region thereof. In some embodiments, a characteristic portion of a polypeptide or region thereof is a portion that is found in the polypeptide or region thereof and in related polypeptide or region thereof that share the particular feature, attribute or activity, but not in those that do not share the particular feature, attribute or activity. In certain embodiments, a characteristic portion shares at least one functional characteristic with the intact polypeptide or region thereof. For example, in some embodiments, a "characteristic portion" of a polypeptide or region thereof is one that contains a continuous stretch of amino acids, or a collection of continuous stretches of amino acids, that together are characteristic of the polypeptide or region thereof. In some embodiments, each such continuous stretch generally contains at least 2, 5, 10, 15, 20, 50, or more amino acids. In general, a characteristic portion of a polypeptide or region thereof is one that, In addtion to the sequence and/or structural Identity specified above, shares at least one functional characteristic with the relevant intact polypeptide or region thereof. In some embodiments, a characteristic portion may be biologically active. In some embodiments, a fragment as described herein can be a portion. Accordingly, in some embodiments, a characteristic fragment can be a "characteristic portion."

[0271] Combination therapy. As used herein, the term "combination therapy" refers to those situations in which a subject is simultaneously exposed to two or more therapeutic regimens (e.g., two or more therapeutic agents (e.g., two or more antibody agents)). In some embodiments, the two or more regimens may be administered simultaneously; in some embodiments, such regimens may be administered sequentially (e.g., all "doses" of a first regimen are administered prior to administration of any doses of a second regimen); In some embodiments, such agents are administered in overlapping dosing regimens. In some embodiments, administration of combination therapy may involve administration of one or more agent(s) or modality(ies) to a subject receiving the other agent(s) or modallty(les) in the combination. For clarity, combination therapy does not require that Individual agents be administered together In a single composition (or even necessarily at the same time), although In some embodiments, two or more agents, or active moieties thereof, may be administered together In a combination composition.

[0272] Comparable; As used herein, the term "comparable" refers to two or more agents, entitles, situations, sets of conditions, etc., that may not be Identical to one another but that are sufficiently slmiar to permit comparison there between so that one skilled In the art will appreciate that conclusions may reasonably be drawn based on differences or similarities observed. In some embodiments, comparable sets of conditions, circumstances, individuals, or populations are characterized by a plurality of substantially identical features and one or a small number of varied features. Those of ordinary skill in the art will understand, in context, what degree of identity is required in any given circumstance for two or more such agents, entities, situations, sets of conditions, etc. to be considered comparable. For example, those of ordinary skill in the art will appreciate that sets of circumstances, individuals, or populations are comparable to one another when characterized by a sufficient number and type of substantially identical features to warrant a reasonable conclusion that differences in results obtained or phenomena observed under or with different sets of circumstances, individuals, or populations are caused by or indicative of the variation in those features that are varied.

[0273] Corresponding toe As used herein, the term "corresponding to" refers to a relationship between two or more entities. For example, the term "corresponding to' may be used to designate the position/identity of a structural element in a compound or composition relative to another compound or composition (e.g., to an appropriate reference compound or composition). For example, in some embodiments, a monomeric residue in a pdymer (e.g., an amino add residue in a polypeptide or a nucleic acid residue in a polynucleotide) may be identified as "corresponding to" a residue in an appropriate reference polymer. For example, those of ordinary skill will appreciate that, for purposes of simplicity, residues in a polypeptide are often designated using a canonical numbering system based on a reference related polypeptide, so that an amino add "corresponding to” a residue at position 190, for example, need not actually be the 190^th amino add in a particular amino add chain but rather corresponds to the residue found at 190 in the reference polypeptide; those of ordinary skill in the art readily appreciate how to identify "corresponding" amino acids. For example, those skilled in the art wil be aware of various sequence alignment strategies, including software programs such as, for example, BLAST, CS-BLAST, CUSASW++, DIAMOND, FASTA, GGSEARCH/GLSEARCH, Genoogle, HMMER, HHpred/HHsearch, IDF, Infernal, KLAST, USEARCH, parasai, PSI-BLAST, PSI-Search, ScalaBLAST, Sequilab, SAM, SSEARCH, SWAPHI, SWAPHI-LS, SWIMM, or SWIPE that can be utilized, for example, to identify "corresponding" residues In polypeptides and/or nudeic acids in accordance with the present disdosure. Those of skill in the art wil also appreciate that, in some instances, the term "corresponding to” may be used to describe an event or entity that shares a relevant similarity with another event or entity (e.g., an appropriate reference event or entity). To give but one example, a gene or protein in one organism may be described as "corresponding to” a gene or protein from another organism in order to indicate, in some embodiments, that it plays an analogous role or performs an analogous function and/or that it shows a particular degree of sequence Identity or homology, or shares a particular characteristic sequence element [0274] Dosing regimen-. Those skilled in the art will appredate that the term "dosing regimen” (or "therapeutic regimen”) may be used to refer to a set of unit doses (typically more than one) that are administered Individually to a subject, typically separated by periods of time. In some embodiments, a given therapeutic agent has a recommended dosing regimen, which may involve one or more doses.

[0275] Encodes, ks used herein, the term "encode" or "encoding" refers to sequence information of a first molecule that guides production of a second molecule having a defined sequence of nucleotides (e.g., a polyribonucleotide) or a defined sequence of amino adds. For example, a DMA molecule can encode an RNA molecule (e.g., by a transcription process that Indudes a DNA-dependent RNA polymerase enzyme). An RNA molecule can encode a polypeptide (e.g., by a translation process). Thus, a gene, a cDNA, or an RNA molecule encodes a polypeptide if transcription and translation of RNA corresponding to that gene produces the polypeptide In a cell or other biological system. In some embodiments, a coding region of a polyribonudeotide encoding a target antigen refers to a coding strand, the nudeotide sequence of which is identical to the polyribonucleotide sequence of such a target antigen. In some embodiments, a coding region of a polyribonudeotide encoding a target antigen refers to a non-coding strand of such a target antigen, which may be used as a template for transcription of a gene or cDNA.

[0276] Expression-. As used herein, the term "expression" of a nucleic acid sequence refers to the generation of a gene product from the nudeic add sequence. In some embodiments, a gene product can be a transcript, e.g., a polyribonucleotide as provided herein. In some embodiments, a gene product can be a polypeptide. In some embodiments, expression of a nudeic add sequence involves one or more of the following: (1) production of an RNA template from a DNA sequence (e.g., by transcription); (2) processing of an RNA transcript (e.g., by splidng, editing, etc.); (3) translation of an RNA into a polypeptide or protein; and/or (4) post-translational modification of a polypeptide or protein. [0277] Heterologous: As used herein, the term "heterologous", with respect to secretory signal or transmembrane region, refers to a secretory signal or transmembrane region from a virus or an organism other than Piasmocfium.

[0278] Homology*. As used herein, the term "homology” or "homolog" refers to the overafl relatedness between polynucleotide molecules (e.g., DNA molecules and/or RNA molecules) and/or between polypeptide molecules. In some embedments, polynucleotide molecules (e.g., DNA molecules and/or RNA mdecides) and/or polypeptide molecules are considered to be "homologous" to one another if their sequences are at least 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% identical. In some embodiments, polynucleotide molecules (e.g., DNA molecules and/or RNA molecules) and/or polypeptide molecules are considered to be "homologous" to one another if their sequences are at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% similar (e.g., containing residues with related chemical properties at corresponding positions). For example, as is well known by those of ordinary skill in the art, certain amino acids are typically classified as similar to one another as "hydrophobic" or "hydrophilic" amino acids, and/or as having "polar' or "non-polar” side chains. Substitution of one amino add for another of the same type may often be considered a "homologous” substitution.

[0279] Identity. As used herein, the term “identity" refers to the overall relatedness between polynucleotide molecules DNA molecules and/or RNA molecules) and/or between polypeptide molecules. In some embodiments, polynucleotide molecules (eg., DNA molecules and/or RNA molecules) and/or between polypeptide molecules are considered to be "substantially Identical" to one another If their sequences are at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% Identical. Calcination of the percent Identity of two nudeic add or polypeptide sequences, for example, can be performed by aligning the two sequences for optimal comparison purposes (e.g., gaps can be introduced in one or both of a first and a second sequence for optimal alignment and non-ldentlcal sequences can be disregarded for comparison purposes). In certain embodiments, the length of a sequence aligned for comparison purposes is at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or substantially 100% of the length of a reference sequence. The nucleotides at corresponding positions are then compared. When a position In the first sequence is occupied by the same residue (e.g., nudeotide or amino add) as the corresponding position in the second sequence, then the molecules are identical at that position. The percent identity between the two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps, and the length of each gap, which needs to be introduced for optimal alignment of the two sequences. The comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm. For example, the percent Identity between two nudeotide sequences can be determined using the algorithm of Meyers and Miller, 1989, which has been incorporated into the ALIGN program (version 2.0). In some exemplary embodiments, nucleic acid sequence comparisons made with the ALIGN program use a PAM120 weight residue table, a gap length penalty of 12 and a gap penalty of 4. The percent identity between two nudeotide sequences can, alternatively, be determined using the GAP program in the GCG software package using an NWSgapdna.CMP matrix.

[0280] Jhcneaswt Induced, or Reduced.* As used herein, these terms or grammatically comparable comparative terms, indicate values that are relative to a comparable reference measurement. For example, in some embodiments, an assessed value achieved with a provided composition (e.g., a pharmaceutical composition) may be "increased" relative to that obtained with a comparable reference composition. Alternatively or additionally, in some embodiments, an assessed value achieved in a subject may be "increased" relative to that obtained in the same subject under different conditions (e.g., prior to or after an event; or presence or absence of an event such as administration of a composition (e.g., a pharmaceutical composition) as described herein, or in a different, comparable subject (e.g., in a comparable subject that differs from the subject of interest in prior exposure to a condition, e.g., absence of administration of a composition (e.g., a pharmaceutical composition) as described herein.). In some embodiments, comparative terms refer to statistically relevant differences (e.g., that are of a prevalence and/or magnitude sufficient to achieve statistical relevance). Those skilled in the art will be aware, or will readiy be able to determine, in a given context, a degree and/or prevalence of difference that is required or sufficient to achieve such statistical significance. In some embodiments, the term "reduced" or equivalent terms refers to a reduction in the level of an assessed value by at least 5%, at least 10%, at least 20%, at least 50%, at least 75% or higher, as compared to a comparable reference. In some embodiments, the term "reduced” or equivalent terms refers to a complete or essentially complete inhibition, i.e., a reduction to zero or essentially to zero. In some embodiments, the term "increased” or "Induced” refers to an increase In the level of an assessed value by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 80%, at least 100%, at least 200%, at least 500%, or higher, as compared to a comparable reference.

[0281] in order: As used herein with reference to a polynucleotide or polyribonucleotide, "In order" refers to the order of features from 5' to 3' along the polynucleotide or polyribonucleotide. As used herein with reference to a polypeptide, "in order” refers to the order of features moving from the N-termlnal-most of the features to the C- termlnal-most of the features along the polypeptide. "In order” does not mean that no additional features can be present among the listed features. For example, If Features A, B, and C of a polynucleotide are described herein as being "In order, Feature A, Feature B, and Feature C,” this description does not exclude, e.g., Feature D being located between Features A and B.

[0282] Isolated. The term "Isolated'' means altered or removed from the natural state. For example, a nucleic add or a peptide naturally present in a living animal is not "Isolated," but the same nucleic add or peptide partiaHy or completely separated from the coexisting materials of Its natural state is "Isolated." An Isolated nucleic add or protein can exist in substantiaRy purified form, or can exist in a non-native environment such as, for example, a host cell. [0283] Linker. As used herein, the term "tinker" refers to a portion of a polypeptide that connects different regions, portions, or antigens to one another.

[0284] Lipid*. As used herein, the terms "lipid" and "lipid-lke material" are broadly defined as molecules which comprise one or more hydrophobic moieties or groups and optionally also one or more hydrophilic moleties or groups. Molecules comprising hydrophobic moieties and hydrophilic moieties are also typically denoted as amphiphiles.

[0285] Merozoite stage spedfic Ptasmodium antigen: As used herein, the term "merozoite stage specific Plasmodium antigen" refers to an antigen that is expressed during the merozoite stage of the Plasmodium life cyde. In some embodiments, a merozoite stage spedfic Plasmodium antigen is a Rh5 invasion complex antigen.

[0286] Multimerization region*. As used herein, the term "multimerization region" refers to a region that directs assembly of multimers into a complex, where each multimer comprises a polypeptide associated with the multimerization region. [0287] RNA lipid nanopartider. As used herein, the term "RNA lipid nanopartfcle* refers to a nanopartide comprising at least one lipid and RNA molecule(s), e.g., one or more polyribonucleotides as provided herein. In some embodiments, an RNA lipid nanopartide comprises at least one cationic amino lipid. In some embodiments, an RNA lipid nanopartide comprises at least one cationic amino lipid, at least one helper lipid, and at least one polymer- conjugated lipid (e.g., PEG-conjugated lipid). In various embodiments, RNA lipid nanoparticles as described herein can have an average size (.e.g., Z-average) of about 100 nm to 1000 nm, or about 200 nm to 900 nm, or about 200 nm to 800 nm, or about 250 nm to about 700 nm. In some embodiments of the present disclosure, RNA lipid nanopartides can have a particle size (e.g., Z-average) of about 30 nm to about 200 nm, or about 30 nm to about 150 nm, about 40 nm to about 150 nm, about 50 nm to about 150 nm, about 60 nm to about 130 nm, about 70 nm to about 110 nm, about 70 nm to about 100 nm, about 80 nm to about 100 nm, about 90 nm to about 100 nm, about 70 to about 90 nm, about 80 nm to about 90 nm, or abort 70 nm to about 80 nm. In some embodiments, an average size of lipid nanoparticles is determined by measuring the average partide diameter. In some embodiments, RNA lipid nanopartides may be prepared by mixing lipids with RNA molecules described herein.

[0288] Neutralization*. As used herein, the term "neutralization" refers to an event in which binding agents such as antibodies bind to a biological active site of a parasite such as a receptor binding protein, thereby inhibiting the parasitic Infedion of cells. In some embodiments, the term "neutralization" refers to an event in which binding agents eliminate or significantly reduce ability of Infecting cells.

[0289] Nucleic add/ Polynucleotide.* As used herein, the term "nucleic add" refers to a polymer of at least 10 nucleotides or more. In some embodiments, a nudek add is or comprises DNA. In some embodiments, a nucleic add is or comprises RNA. In some embodiments, a nucleic add Is or comprises peptide nudek add (PNA). In some embodiments, a nudek acid is or comprises a single stranded nudek add. In some embodiments, a nucleic acid is or comprises a double-stranded nucleic add. In some embodiments, a nudek acid comprises both single and doublestranded portions. In some embodiments, a nudek add comprises a backbone that comprises one or more phosphodiester linkages. In some embodiments, a nudek add comprises a backbone that comprises both phosphodiester and non-phosphodlester linkages. For example, in some embodiments, a nudek add may comprise a backbone that comprises one or more phosphorothtoate or S'-N-phosphoramldlte linkages and/or one or more peptide bonds, e.g., as in a "peptide nudek add". In some embedments, a nudek add comprises one or more, or all, natural residues (e.g., adenine, cytosine, deoxyadenosine, deoxycytidine, deoxyguanosine, deoxythymidine, guanine, thymine, uradl). In some embodiments, a nudek add comprises on or more, or all, non-natural residues. In some embodiments, a non-natural residue comprises a nucleoside analog (e.g., 2-aminoadenosine, 2- thiothymidine, inosine, pyrrolo-pyrimidine, 3 -methyl adenosine, 5-methykytidine, C-5 propynyl-cytidine, C-5 propynyl-uridine, 2-aminoadenosine, C5-bromouridlne, C5-fluorouridfrie, C5-lodouridlne, C5-propynyl-uridlne, C5 - propynyl-cytidine, C5-methylcytidine, 2-aminoadenosine, 7-deazaadenosine, 7-deazaguanosine, 8-oxoadenosine, 8- oxoguanosine, 6-O-methylguanine, 2-thiocytidine, methylated bases, intercalated bases, and combinations thereof). In some embodiments, a non-natural residue comprises one or more modified sugars (e.g., 2'-fluororibose, ribose, 2'-deoxyribose, arabinose, and hexose) as compared to those in natural residues. In some embodiments, a nudek add has a nucleotide sequence that encodes a functional gene product such as an RNA or polypeptide. In some embodiments, a nudek acid has a nucleotide sequence that comprises one or more introns. In some embodiments, a nucleic add may be prepared by isolation from a natural source, enzymatic synthesis (e.g., by polymerization based on a complementary template, e.g., in vivo or in vitro), reproduction in a recombinant cell or system, or chemkal synthesis. In some embodiments, a nucleic acid is at least 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 600, 700, 800, 900, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, 5000, 5500, 6000, 6500, 7000, 7500, 8000, 8500, 9000, 9500, 10,000, 10,500, 11,000, 11,500, 12,000, 12,500, 13,000, 13,500, 14,000, 14,500, 15,000, 15,500, 16,000, 16,500, 17,000, 17,500, 18,000, 18,500, 19,000, 19,500, or 20,000 or more residues or nucleotides long.

[0290] Pharmaceutically effective amount The term "pharmaceutically effective amount" or

“therapeutically effective amount" refers to the amount which achieves a desired reaction or a desired effect alone or together with further doses. In the case of the treatment of a particular disease (e.g., malaria), a desired reaction In some embodiments relates to Inhibition of the course of the disease (e.g., malaria). In some embodiments, such inhibition may comprise slowing down the progress of a disease (e.g., malaria) and/or interrupting or reversing the progress of the disease (e.g., malaria). In some embodiments, a desired reaction in a treatment of a disease (e.g., malaria) may be or comprise delay or prevention of the onset of a disease (e.g., malaria) or a condition (e.g., a malaria associated condition). An effective amount of a composition (e.g., a pharmaceutical composition) described herein will depend, for example, on disease (e.g., medaria) or a condition (e.g., a malaria associated condition) to be treated, the severity of such a disease (e.g., malaria) or a condition (e.g., a malaria associated condition), Individual parameters of the patient, Including, e.g., age, physiological condition, size and weight, the duration of treatment, the type of an accompanying therapy (if present), the specific route of administration and similar factors.

Accordingly, doses of a composition (e.g., a pharmaceutical composition) described herein may depend on various of such parameters. In the case that a reaction In a patient is Insufficient with an initial dose, higher doses (or effectively higher doses achieved by a different, more localized route of administration) may be used.

[0291] Polypeptide.- As used herein, the term “polypeptide' refers to a polymeric chain of amino adds. In some embodiments, a polypeptide has an amino acid sequence that occurs In nature. In some embodiments, a polypeptide has an amino add sequence that does not occur In nature. In some embodiments, a polypeptide has an amino acid sequence that is engineered In that It is designed and/or produced through action of the hand of man. In some embodiments, a polypeptide may comprise or consist of natural amino adds, non-natural amino acids, or both. In some embodiments, a polypeptide may comprise or consist of only natural amino adds or only non-natural amino adds. In some embodiments, a polypeptide may comprise D-amino adds, L-amino acids, or both. In some embodiments, a polypeptide may comprise only D-amino adds. In some embodiments, a polypeptide may comprise only L-amino acids. In some embodiments, a polypeptide may indude one or more pendant groups or other modifications, e.g., modifying or attached to one or more amino add side chains, at the polypeptide's N-terminus, at the polypeptide's C-termlnus, or any combination thereof. In some embodiments, such pendant groups or modifications comprise acetylation, amidation, lipidation, methylation, pegylation, etc., including combinations thereof. In some embodiments, a polypeptide may be cydic, and/or may comprise a cydic portion. In some embodiments, a polypeptide is not cyclic and/or does not comprise any cydic portion. In some embodiments, a polypeptide is linear. In some embodiments, a polypeptide may be or comprise a stapled polypeptide. In some embodiments, the term "polypeptide" may be appended to a name of a reference polypeptide, activity, or structure; in such instances it is used herein to refer to polypeptides that share the relevant activity or structure and thus can be considered to be members of the same class or family of polypeptides. For each such class, the present specification provides and/or those skilled in the art will be aware of exemplary polypeptides within the dass whose amino acid sequences and/or functions are known; in some embodiments, such exemplary polypeptides are reference polypeptides for the polypeptide class or family, in some embodiments, a member of a polypeptide class or family shows significant sequence homology or identity with, shares a common sequence motif (e.g., a characteristic sequence element) with, and/or shares a common activity (in some embodiments at a comparable level or within a designated range) with a reference polypeptide of the dass; in some embodiments with all polypeptides within the class). For example, in some embodiments, a member polypeptide shows an overall degree of sequence homology or identity with a reference polypeptide that is at least about 30-40%, and is often greater than about 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more and/or includes at least one region (e.g., a conserved region that may in some embodiments be or comprise a characteristic sequence element) that shows very high sequence identity, often greater than 90% or even 95%, 96%, 97%, 98%, or 99%. Such a conserved region usually encompasses at least 3-4 and often up to 35 or more amino adds; in some embodiments, a conserved region encompasses at least one stretch of at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 or more contiguous amino adds. In some embodiments, a relevant polypeptide may comprise or consist of a fragment of a parent polypeptide. In some embodiments, a polypeptide is a Plasmodium polypeptide construct described herein. A Plasmodium polypeptide construct is a polypeptide that indudes one or more malarial proteins, or one or more portions thereof. In some embodiments, a Plasmodium polypeptide construct described herein Indudes at least one region of Plasmodium Rh5 Invasion complex polypeptide or an antigenic a portion thereof. In some embodiments, a Plasmodium polypeptide construct additionally includes one or more additional amino add sequences, such as a secretory signal (e.g., a heterologous secretory signal), a transmembrane region (e.g., a heterologous transmembrane region), a multlmerization region, and/or a linker, as described herein.

[0292] Prevent As used herein, the term "prevent" or "prevention" when used In connection with the occurrence of a disease, disorder, and/or condition, refers to reducing the risk of developing the disease, disorder and/or condition and/or to delaying onset of one or more characteristics or symptoms of the disease, disorder or condition. Prevention may be considered complete when onset of a disease, disorder or condition has been delayed for a predefined period of time. In some embodiments, prevention refers to reducing the risk of developing clinical malaria.

[0293] Referencer. As used herein, the term "reference" describes a standard or control relative to which a comparison is performed. For example, in some embodiments, an agent, animal, individual, population, sample, sequence or value of interest is compared with a reference or control agent, animal, individual, population, sample, sequence or value. In some embodiments, a reference or control is tested and/or determined substantially simultaneously with the testing or determination of Interest In some embodiments, a reference or control Is a historical reference or control, optionally embodied in a tangible metfium. Typically, as would be understood by those skilled in the art, a reference or control is determined or characterized under comparable conditions or circumstances to those under assessment Those skilled in the art will appreciate when sufficient similarities are present to justify reliance on and/or comparison to a particular possible reference or control.

[0294] Rh5 invasion complex-. As used herein refers to a complex that is present in Plasmodium merozoites that includes Rh5. In some embodiments, a Rh5 invasion complex facilitates merozoite binding to and/or invasion of an erythrocyte. Without limitation, polypeptides in an Rh5 invasion complex may include one or more of Plasmodium reticulocyte -binding protein homolog 5 (Rh5), Plasmodium Cysteine-Rich Protective Antigen (CyRPA), Plasmodium Rh5-interacting Protein (Ripr), Plasmodium Pl 13, Absmod't/m thrombospondin-related apical merozoite protein (TRAMP), and Plasmodium cysteine-rich small secreted protein (CSS). Exemplary Rh5 invasion complexes include the RCR complex and the PCRCR complex. An RCR complex is a trimeric complex comprising Rh5, cysteine-rich protective antigen (CyRPA), and Rh5-interacting protein (Ripr). A PCRCR complex is a pentameric complex comprising Rh5, cysteine-rich small secreted (CSS), Ripr, CyRPA, and Piasmodum thrombospondin-related apical merozoite protein (PTRAMP).

[0295] RibonucMc add (RNA) or Polyribonucleotide: As used herein, the term "ribonucleic add," "RNA,” or "polyribonudeotide" refers to a polymer of ribonucleotides. In some embodiments, an RNA is single stranded. In some embodiments, an RNA Is double stranded. In some embodiments, an RNA comprises both single and double stranded portions. In some embodiments, an RNA can comprise a backbone structure as described In the definition of "Nudeic add / Polynudeotide” above. An RNA can be a regulatory RNA (e.g., siRNA, microRNA, etc), or a messenger RNA (mRNA). In some embodiments, an RNA is a mRNA. In some embodiments, where an RNA is a mRNA, a RNA typically comprises at its 3' end a poly(A) region. In some embodiments, where an RNA is a mRNA, an RNA typically comprises at its 5* end an art-recognized cap structure, e.g., for recognizing and attachment of a mRNA to a ribosome to initiate translation. In some embodiments, a RNA is a synthetic RNA. Synthetic RNAs indude RNAs that are synthesized rh vitro (e.g., by enzymatic synthesis methods and/or by chemical synthesis methods). In some embodiments, a polyribonucleotide encodes a polypeptide, which Is preferably Is a Plasmodium polypeptide construct

[0296] Ribonudeotide: As used herein, the term ’ribonucleotide" encompasses unmodified ribonudeotldes and modified ribonudeotldes. For example, unmodified ribonudeotldes Indude the purine bases adenine (A) and guanine (G), and the pyrimidine bases cytosine (C) and uracil (U). Modified ribonucleotides may include one or more modifications Including, but not limited to, for example, (a) end modifications, e.g., 5' end modifications (e.g., phosphorylation, dephosphorylation, conjugation, Inverted linkages, etc.), 3' end modifications (e.g., conjugation, inverted linkages, etc.), (b) base modifications, e.g. , replacement with modified bases, stabilizing bases, destabilizing bases, or bases that base pair with an expanded repertoire of partners, or conjugated bases, (c) sugar modifications (e.g., at the 2' position or 4' position) or replacement of the sugar, and (d) intemudeoside linkage modifications, Including modification or replacement of the phosphodiester linkages. The term "ribonucleotide" also encompasses ribonucleotide triphosphates including modified and non-modified ribonucleotide triphosphates. [0297] Secretory signal: &s used herein, the term "secretory signal" refers to an amino add sequence motif that targets associated polypeptides for translocation to a secretory pathway.

[0298] Subject As used herein, the term "subject" refers to an organism to be administered with a composition described herein, e.g., for experimental, diagnostic, prophylactic, and/or therapeutic purposes. Typical subjects indude animals (e.g., mammals such as mice, rats, rabbits, non-human primates, domestic pets, etc.) and humans. In preferred embodiments, a subject is a human subject. In some embodiments, a subject is suffering from a disease, disorder, or condition (e.g., malaria and/or a malaria-associated condition). In some embodiments, a subject is susceptible to a disease, disorder, or condition (e.g., malaria and/or a malaria-assodated condition). In some embodiments, a subject displays one or more symptoms or characteristics of a disease, disorder, or condition (e.g., malaria and/or a malaria-associated condition). In some embodiments, a subject displays one or more nonspecific symptoms of a disease, disorder, or condition (e.g., malaria and/or a malaria-assodated condition). In some embodiments, a subject does not display any symptom or characteristic of a disease, disorder, or condition (e.g., malaria and/or a malaria-assodated condition). In some embodiments, a subject is someone with one or more features characteristic of susceptibility to or risk of a disease, disorder, or condition (e.g., malaria and/or a malaria- assodated condition). In some embodiments, a subject is a patient. In some embodiments, a subject is an individual to whom diagnosis and/or therapy is and/or has been administered.

[0299] Suffering from*. An indvidual who is "suffering from” a disease, disorder, and/or condition (e.g., malaria and/or a malaria-associated condition) has been diagnosed with and/or displays one or more symptoms of a disease, disorder, and/or condition.

[0300] Susceptible te An individual who is "susceptible to" a disease, disorder, and/or condition (e.g., malaria and/or a malaria-associated condition) is one who has a higher risk of developing the disease, disorder, and/or condition (e.g., malaria and/or a malaria-associated condition ) than does a member of the general public. In some embodiments, an individual who is susceptible to a disease, disorder and/or condition (e.g., malaria and/or a malaria-associated condition) may not have been diagnosed with the disease, disorder, and/or condition (e.g., malaria and/or a malaria-associated condition). In some embodiments, an individual who is susceptible to a disease, disorder, and/or condition (e.g., malaria and/or a malaria-associated condition) may exhibit symptoms of the disease, disorder, and/or condition (e.g., malaria and/or a malaria-associated condition). In some embodiments, an individual who is susceptible to a disease, disorder, and/or condition (e.g., malaria and/or a malaria-associated condition) may not exhibit symptoms of the disease, disorder, and/or condition (e.g., malaria and/or a malaria-assodated condition). In some embodiments, an Individual who is susceptible to a disease, disorder, and/or condttion (e.g., malaria and/or a malaria-assodated condition) will develop the disease, disorder, and/or condition (e.g., malaria and/or a malaria- assodated condition). In some embodiments, an Individual who is susceptible to a disease, disorder, and/or condition (e.g., malaria and/or a malaria-assodated condition) will not develop the disease, disorder, and/or condition (e.g., malaria and/or a malaria-associated condition).

[0301] Therapy. The term "therapy" refers to an administration or delivery of an agent or intervention that has a therapeutic effect and/or elicits a desired biological and/or pharmacological effect (e.g., has been demonstrated to be statistically likely to have such effect when administered to a relevant population). In some embodiments, a therapeutic agent or therapy is any substance that can be used to alleviate, ameliorate, relieve, inhibit, prevent, delay onset of, reduce severity of, and/or reduce incidence of one or more symptoms or features of a disease, disorder, and/or condition (e.g., malaria and/or a malaria-assodated condition). In some embodiments, a therapeutic agent or therapy is a medical intervention that can be performed to alleviate, relieve, inhibit, present, delay onset of, reduce severity of, and/or reduce incidence of one or more symptoms or features of a disease, disorder, and/or condition.

[0302] Transmembrane region; As used herein, the term "transmembrane region" refers to a region of a polypeptide that spans a biological membrane, such as the plasma membrane of a cell.

[0303] Theati As used herein, the term "treat,” "treatment," or "treating" refers to any method used to partially or completely alleviate, ameliorate, relieve, inhibit, prevent, delay onset of, reduce severity of, and/or reduce inddence of one or more symptoms or features of a disease, disorder, and/or condition (e.g., malaria and/or a malaria-associated condition). Treatment may be administered to a subject who does not exhibit signs of a disease, disorder, and/or condition (e.g., malaria and/or a malaria-assodated condition). In some embodiments, treatment may be administered to a subject who exhibits only early signs of the disease, disorder, and/or condition (e.g., malaria and/or a malaria-assodated condition), for example for the purpose of decreasing the risk of developing pathology associated with the disease, disorder, and/or condition. In some embodiments, treatment may be administered to a subject at a later-stage of disease, disorder, and/or condition (e.g., malaria and/or a malaria- assodated condition).

[0304] Variant: As used herein, the term "variant" refers to a molecule that shows significant structural (e.g., primary or secondary) identity with a reference molecule but differs structurally from the reference molecule. For example, a variant polypeptide or nucleic add may differ from a reference polypeptide or nudeic add as a result of one or more differences in amino acid or nucleotide sequence and/or one or more differences in chemical moieties (e.g., carbohydrates, lipids, phosphate groups) that are covalently components of the polypeptide or nudeic add (e.g_v that are attached to the polypeptide or nudeic acid backbone).

DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS

Malaria

[0305] Malaria is a mosquito-borne infectious cfisease caused by single-celled eukaryotic Plasmodium parasites that are transmitted by the bite of Anopheles app. mosquitoes (Phillips, M., etai. Malaria. Nat Rev Dis Primers3, 17050, 2017, which is incorporated herein by reference in its entirety). Mosquitoes that transmit malaria must have been infected through a previous blood meal taken from an infected subject (e.g., a human). When a mosquito bites an infected subject a small amount of blood is taken in containing Malaria parasites. The infected mosquito can then subsequently bite a non-infected subject, infecting the subject [0306] Malaria remains one of the most serious infectious diseases, causing approximately 200 million dinical cases and 500,000-600,000 deaths annuaHy. Although significant effort has been invested in developing therapeutic treatments for malaria, many malaria parasites have developed resistance to available therapeutics. According to Malaria Eradication Research Agenda Initiative, malaria eradication will only be achievable through effective vaccination.

[0307] In 2015, the European Medicines Agency gave a positive review to a malaria vaccine candidate known as *RTS,S", a milestone in malaria vaccine development. In 2019, the World Health Organization launched pilot programs that provide RTS,S to children at least 5 months of age in parts of three sub-Saharan African countries. RT5,S/AS01 Is an adjuvanted protein subunit vaccine that consists of a portion of the major repeat region and the C- terminus of CSP from Plasmodium falciparum fused to the Hepatitis B surface antigen (HBsAg). The vaccine is a mix of this ACSP-HBsAg compound with HBsAg that forms virus-like particles (RTS,S/AS01; Mosquirix™). RTS,S is administered according to a regimen that requires four doses: an initial 3-dose schediie given at least 1 month apart, and a 4th dose 15-18 months after dose 3 (see, for example, Vandoolaeghe & Schuerman Expert Rev Vaccines. 15:1481, 2016; PATH_MVI_RTSS_Fact Sheet_042019, each of which Is incorporated herein by reference In its entirety). Reports indicate that RTS,S protects approximately 30% to 50% of children from clinical disease over 18 months. RTS,S has been reported to induce protective antibody and CD4+ T-cell responses, but only negligible CD8+ T cell responses (see, for example, Moris et al. Hum Vaccki Immunother 14:17, 2018, which is incorporated herein by reference in Its entirety). Phase III studies of RTS,S delivered as a three-dose series with a booster after 1 yr (year) showed moderate vaccine efficacy in children aged 5 to 17 months preventing 36% of clinical malaria cases over the ful study period with a median follow-up of 4 yrs, with a range of 20% in high to 66% In low transmission settings. Furthermore, published literature suggests that protection wanes over time InducHng reports of potential negative efficacy after 5 yrs In children with high malaria exposure (Olotu et al. 2016, N. Engl. J. Med. 374:2519-29) which is incorporated herein by reference in its entirety).Thus, an effective malaria vaccine remains an unmet medical need of critical importance for global health.

Lifecycle

[0308] During a blood meal, infected mosquitos inject, along with their anticoagulating saliva, sporozoites known as the liver stage of Plasmodium spp. Sporozoites journey through the skin, and if successful, into hepatocytes of the liver. This journey happens very quickly; it can be completed within only a few minutes (Sinnis et al., Parasite! Int. 2007 Sep;56(3): 171-8, which is incorporated herein by reference in its entirety). This is a time known to be a stage during which it is challenging for the host immune system to mount an effective response against the parasite, as only a small number (thought to be a few hundred at maximum) of sporozoites are injected by the mosquito, with a small fraction of those parasites establishing Infection In the liver (Flores-Garda et al., mBio. 2018 Nov 20;9(6):e02194-18, which is incorporated herein by reference in its entirety).

[0309] When moving from an inoculation site in the skin to the liver, sporozoites traverse host ceHs (Mota et al., Science 2001 Jan 5;291(5501): 141-4). Sporozoites traverse different types of host cells at the dermis, including fibroblasts and phagocytes (Amino et al.. Cell Host Microbe. 2008 Feb 14;3(2):88-96, which is incorporated herein by reference in its entirety), and the liver sinusoidal barrier, containing liver endothelial cells and Kupffer cells (Frevert et al., PLoS Biol 3(6): el92. 2005, which is incorporated herein by reference In its entirety) and sinusoidal endothelial cells (Tavares et al., J Exp Med 2013 May 6;210(5):905-15, which is Incorporated herein by reference In Its entirety), in order to gain access to hepatocytes. Sporozoites preferentially traverse cells with low-sulfated heparin sulfate proteoglycans (HSPGs) but preferentially Invade cells with high-sulfated HSPGs (Coppi et al., Cell Host & Microbe 2, 316-327, November 2007, which Is Incorporated herein by reference In Its entirety).

[0310] Cell traversal was first observed as non-phagocytic entry of P. Aergite/sporozoltes Into macrophages followed by "escape" from these cels (Vanderberg et al., J. Euk. Microbiol. 37:528-536, 1990, which is Incorporated herein by reference In Its entirety). The biochemical, biophysical, and stepwise processes of traversal are still being explored. However, It has been suggested by electron microscopy that host cell rupture occurs upon entry and exit from the host cell (Mota et al., 2001; Tavares et al., 2013, each of which is incorporated herein by reference in its entirety). It has also been shown that P. yoetiispotozoktes can enter hepatocytes via a transient vacuole and that host membrane rupture occurs upon cell exit rather than cell entry (Risco-Castillo et al., Cell Host Microbe 2015 Nov ll;18(5):593-603, which is incorporated herein by reference in its entirety).

[0311] Sporozoites also traverse hepatocytes before establishing a productive hepatocyte infection (Mota et al., 2001, which is incorporated herein by reference in its entirety). Several possibilities emerged as to why this occurs. The first hypothesis suggested that migration through hepatocytes primes parasites for invasion by activating apical exocytosis (Mota et al., Nat Med 2002 Nov;8(ll): 1318-22, which is incorporated herein by reference in Its entirety). The second theory suggested that traversal releases hepatocyte growth factor (HGF), making neighboring hepatocytes more susceptible to infection (Carrolo et al., Nat Med . 2003 Nov;9(ll):1363-9, which is incorporated herein by reference in its entirety). Lastly, other studies suggest that it takes some time for sporozoites to switch off the machinery for traversal and activate invasion machinery (Amino et al., 2008, Coppi et al., 2007, each of which is incorporated herein by reference in its entirety), and that traversal primarily functions to penetrate cell banners and avoid phagocytosis en route to the liver (Amino et al., 2008, Coppi et al., 2007, Tavares et al., 2013, each of which is incorporated herein by reference in its entirety). [0312] Although it has been shown that sporozoites traverse human cells (Behet et al., Malar J 2014 Apr 5;13: 136; Cha et al., J Exp Med 2015 Aug 24;212(9):139M03; Dumoulin et al., PLoS One 2015 Jun 12;10(6):e0129623; van Schaijk et al., PLoS ONE, 3 (10). e3549 2008, each of which is incorporated herein by reference in its entirety), the molecular basis for the traversal process is largely unstudied. Antibodies against circumsporozoite protein (CSP) impair traversal (Dumoulin et al., 2015, which is incorporated herein by reference in its entirety), but this is likely due to inhibition of motility rather than a direct effect (Cha et al., J Exp Med 2016 Sep 19;213(10):2099-112, which is incorporated herein by reference in its entirety). Furthermore, antibodies induced by CPS-immunication (chloroquine prophylaxis with sporozoites) may target CSP and interfere with cell traversal (Behet et al., 2014, which is Incorporated herein by reference in its entirety). Recently it was shown that glyceraldehyde 3- phosphate dehydrogenase (GAPDH) on the parasite surface Interacts with CD68 on Kupffer cells during traversal (Cha et al., 2015, Cha et al., 2016, each of which is incorporated herein by reference in its entirety).

[0313] In rodent malaria parasites such as P. berghei, two sporozoite miaroneme proteins have been identified that appear to be essential for cel traversal (sporozoite microneme protein essential for cell traversal [SPECT1; Ishino et al., PLoS Biol., 2 (2004), pp. 77-84, which is incorporated herein by reference in its entirety] and SPECT2 [Ishino et al., Cell. Microbiol., 7 (2005), pp. 199-208, which is incorporated herein by reference in its entirety], also called perforin-like protein 1 [PLP1] [Kaiser et al., Mol. Blochem. Parasitol., 133 (2004), pp. 15-26, which is Incorporated herein by reference In Its entirety]. Even though genetic disruption of SPECT1 or SPECT2 rendered sporozoites unable to traverse murine cells, they still invaded hepatocytes in vitro (Ishino et al., 2004, Ishino et al., 2005, each of which Is Incorporated herein by reference in its entirety). When Injected Into rodents, sporozoites lacking SPECT1 or SPECT2 were Impaired for liver infection, but a small number of sporozoites could stIH establish liver infection that resulted in subsequent patency. However, depletion of Kupffer cells aflowed mutants to establish liver infection at levels comparable with wild-type parasites (Ishino et al., 2004, Ishino et al., 2005, each of which Is incorporated herein by reference In its entirety). This data suggests that traversal by rodent-infecting sporozoites Is Important for navigating through the sinusoidal layer, but not for hepatocyte invasion, malarial exoerythrocytic forms development, or growth within erythrocytes (Ishino et al., 2004, Ishino et al., 2005, each of which is Incorporated herein by reference In its entirety).

[0314] The ortholog of SPECT2 in P. yoeSi, PLP1, has been shown to play a role in cell traversal. Although this protein is not required for hepatocyte entry, it plays a role in egress from transient vacuoles during traversal (Risco- Castillo et al., 2015, each of which is incorporated herein by reference in its entirety). Thus, sporozoites that infect rodents can traverse host cells by generating a vacuole at the entry step and use a perforin-like protein (e.g., SPECT2/PLP1) to escape from this compartment and/or a host cell, during cell exit.

[0315] Once sporozoites have Invaded liver cells, they dfferentiate into merozoites, a replicative form of the parasite. Within a few days, a single sporozoite can lead to 5,000-10,000 merozoites. Merozoites bud from the host hepatocyte in structures called merosomes, which contain up to a thousand merozoites and are hidden from host immune responses due to their host membrane composition. These merosomes ultimately release merozoites that mix freely with the host cytoplasm and appear in the blood stream, where they invade red blood cells and begin the blood stage of infection, characterized by fast asexual replication and clinical presentation. Within a small number of days, hundreds of thousands of parasites can be present in the blood.

[0316] Plasmodium spp. parasites gain entry into red blood cells through specific ligand-receptor interactions mediated by proteins on the surface of the parasite that interact with receptors on the host erythrocyte (mature red blood cell) or reticulocyte (immature red blood cell), whereas P. /atoparumcan invade and replicate in erythrocytes and reticulocytes, P. wVaxand other species predominantly invade reticulocytes, which are less abundant than erythrocytes. Most of the erythrocyte-binding proteins or reticulocyte-binding proteins that have been associated with invasion are redundant or are expressed as a family of variant forms; however, for P. falciparum, two essential red blood cell receptors (basigin and complement decay-accelerating factor (also known as CD55)) have been identified.

[0317] P. mar and P. ovale can also enter a dormant state in the liver, the hypnozoite.

[0318] The invasion of a red blood cell by the merozoite involves interactions of multiple parasite derived proteins (e.g., ligands) with red blood cell (RBC) proteins (e.g., receptors) (Weiss et al., Pios Path., 2015 Feb 27; 10.1371, which is Incorporated herein by reference in its entirety). The invasion process begins by the merozoite first attaching to the RBC. Merozoite-RBC Interaction Is further strengthened through merozoite surface protein- 1 (MSP1) and unknown RBC proteins, causing some deformation of the RBC surface. After the deformation, interaction of merozoite EBA and Rh protein families (excluding Rh5) with host receptors like CR1 lead to actin-dependent deformation of the RBC membrane and reorientation of the merozoites apical end onto the RBC surface (Geoghehan et al., Nat Commui., 2021 June 15;10.1038 which is incorporated herein by reference in its entirety). This deformation event increases the surface area of interaction between merozoite and RBC membranes, an interaction which can be stabilized by an Rh5 invasion complex comprising Plasmodium thrombospondin-related apical merozoite protein (PTRAMP), cystelne-rlch small secreted (CSS), cysteine-rich protective antigen (CyRPA), Rh5-interactlng protein (Rlpr) and Rh5. This Rh5 invasion complex including the five proteins is referred to as the PCRCR complex. [0319] The creation of the PCRCR complex begins in the endoplasmic reticulum, where Its constituents cystelne-rlch small secreted (CSS) protein and Plasmodium thrombospondin-related apical merozoite protein (PTRAMP) Interact to form a heterodimer referred to as PTRAMP-CSS. PTRAMP-CSS Is trafficked to a secretory organelle called the microneme, where CSS interacts with Rh5-interactlng protein (Rlpr) and cystelne-rlch protective antigen (CyRPA) to form a tetrameric complex called PCRC. At some point after deformation occurs, the merozoite Initiates a polarized secretion process that allows the mlcronemal PCRC complex opportunity to Interact with rhoptry protein Rh5, forming the pentameric complex PCRCR. The PCRCR complex, through Rh5, can then bind a host cell receptor, called basigin, broadly across the merozoite-RBC Interphase. PCRCR binding to basigin creates a stable and Irreversible platform between the apical end of the merozoite and the deformed RBC surface, allowing Rh5 and Rlpr to embed into the RBC membrane (Scally et al., Nat Microb., 2022 May 4, which is incorporated herein by reference in its entirety). Once the merozoite-RBC interaction is secured, an open connection between the merozoite's apical tip and the RBC surface is formed which acts as a conduit for Ca²* to flow into the RBC, and for merozoite derived invasion proteins, such as AMA1 and RON2, to help establish the moving junction (Srinivasan et al., Proc Natl Acad Sd., 2011 Jul 25;13275-80, which is Incorporated herein by reference in its entirety). It Is through this moving junction that the parasite, utilizing its actin-myosin dependent gliding motility, propels itself inside the RBC and establishes a parasitopherous vacuole within which the parasite grows and replicates.

[0320] It has been shown that Rh5 binds basigin with higher affinity when complexed with CyRPA and Ripr to form the RCR complex, and has the highest affinity for basigin when part of the PCRCR complex (Wong et al., Nature., 2019 and Scally et al., Nat Microb., 2022 May 4, each of which are incorporated herein by reference in its entirety).

[0321] Merozoites that egress from red blood cells can invade other red blood cells to continue to the asexual blood stage of the parasite lifecycle. A small percentage of schizonts are already committed to a different fate and merozoites from these schizonts, following invasion of a new red blood cell, will differentiate into the sexual forms on the parasite's life cycle; either male or female, gametocytes. Gametocytes are taken up by the mosquito vector during blood feeding. In the gut of the mosquito, male gametocytes undergo 3 rapid rounds of mitosis to form 8 flagellate microgametes. Female gametocytes mature into macrogametes, egressing from their red blood cell. Male microgametes are motile forms with flagellae and seek the female macrogamete. The male and female gametocytes fuse, forming a diploid zygote, which matures and differentiates into an ookinete; this motile form secretes chitinases and other lytic proteins such as CelTOS in order to disrupt and traverse through the peritrophic matrix and traverse the midgut epithelium to reach the basal lamina where it further differentiates and matures as an oocyst Oocysts mature over approximately 10 days (depending on the temperature), replicating to form sporozoites that egress the mature oocyst Into the hemocoel of the mosquito. Thousands of sporozoites form in a single oocyst and become randomly distributed throughout the hemocoel. It is thought that sporozoites are passively circulated through the mosquito haemolymph until they encounter the salivary glands, where they actively invade the glands. Following invasion of the salivary gland, sporozoites are re-programmed via an unknown mechanism to prepare for Iver Invasion. Evidence of this reprogramming has been demonstrated by the inability of midgut sporozoites (directly from oocysts) to invade hepatocytes, and also by the fact that sporozoites which have successfully invaded a salivary gland are unable to re-invade another salivary gland if presented one. Salivary gland sporozoites alter mosquito behavior and salivary gland function, as less saliva is produced resulting In an Increase In mosquito probing behavior, increasing the chances of transmission to a human host via a mosquito bite and continuing the human host's phase of this parasite's life cyde.

[0322] P. vivax anti P. ovale can also enter a dormant state in the liver, the hypnozoite. [0323] One particular challenge associated with mounting an immune response capable of clearing sporozoites before a liver infection has been established Is that the most abundant and Immunogenic protein on the sporozoite surface, the circumsporozoite protein (CSP), is only exposed to the immune system in small quantities and for a short duration of time due to the variably low inoculum from the mosquito and the kinetics of hepatocyte infection after inoculation. Furthermore, after liver infection is established, the parasite differentiates into a stage during which it no longer expresses CSP and hstead has a different mosaic of surface antigens, making the parasite Invisible to, e.g., any memory immune response directed towards the liver stage. The early stages of parasite exposure to the human host create a short window for the innate and memory immune systems to generate a response that is capable of clearing all parasites before infection is established. The clinical significance of immune evasion by sporozoites are high at this point, as all it takes is a single sporozoite to invade a hepatocyte and produce as many as 10,000 blood stage parasites (merozoites) capable of invading, replicating within, and lysing red blood cells (Shears et al., jproteome. 2019 July 23; 3404-3418, which Is Incorporated herein by reference In its entirety). These initial stages of the parasite's life cyde in a human host help to demonstrate the difficulty in targeting liver stage parasites for therapeutic intervention and the low efficacy seen with current vaccines. (Oluta A. et al., N Engl J Med. 2016 June 30;216:374, which is incorporated herein by reference in its entirety). Therefore, targeting the blood stage of Plasmodium spp. infection offers a promising approach for reducing deaths caused by malaria.

[0324] Malaria symptoms typically develop 4-8 days after initial red blood cell invasion. Replication cyde of merozoites within the red blood cells continues for 36-72 hours, until hemolysis, releasing the merozoites for another round of red blood cell infection. Thus, in synchronous infections (infections that originate from a single infectious bite), fever occurs every 36-72 hours, when infected red blood cells lyse and release endotoxins en masse. [0325] Some drugs that prevent Plasmodium spp. invasion or proliferation in the liver have prophylactic activity, drugs that block the red blood cell stage are required for the treatment of the symptomatic phase of the disease, and compounds that inhibit the formation of gametocytes or their development in the mosquito (including drugs that kill mosquitoes feeding on blood) are transmission-blocking agents (Phillips, et al. Malaria. Nat Rev Dis Primers3, 17050 (2017), which is incorporated herein by reference in its entirety). B. Genome

[0326] Since completion of the first sequence of P. falciparum 3D7 genome in 2002, genomic research on malaria parasites has rapidly advanced. Except for a short diploid phase after fertilization in the mosquito midgut, Plasmodium spp. parasites are haploid throughout their life cycle. The genomes of (Afferent species range from 20 to 35 megabases, contain 14 chromosomes, a circular plastid genome of approximately 35 kilobases, and multiple copies of a 6 kilobase mitochondrial DMA. Comparison of genomes from different species showed that homologous genes are often found in synthetic blocks arranged in different orders among different chromosomes.

[0327] The adenine-thymine (AT) content of Plasmodium spp. can also be very different, e.g., ~80% AT in P. falciparum, P. reichenowi, and P. gaHinaceunr, ~75% AT in rodent malaria parasites; and ~60% AT in P. vivax, P. knowtesi, and P. cynomolgi. AT content is often higher in introns and intergenic noncoding regions than in proteincoding exons, with an average of 80.6% AT for the whole P. fakiparum genome versus 86.5% for noncoding sequences. The high AT content of P. fakiparum reflects large numbers of low-complexity regions, simple sequence repeats, and microsatellites, as well as a highly skewed codon usage bias. Polymorphisms of AT-rich repeats provide abundant markers for linkage mapping of drug resistance genes and for tracing the evolution and structure of parasite populations.

[0328] Malaria parasite genomes carry multigene families that serve important roles in parasite interactions with their hosts, including, for example, antigenic variation, signaling, protein trafficking, and adhesion. Among the gene families, genes encoding P. fakiparum erythrocyte membrane protein 1 (P/EMP1) have been studied most extensively. Each Individual P. falciparum parasite carries a unique set of 50 to 150 copies of the vargene In Its genome, where switches of gene expression can produce antigenic variation. P/EMP1 plays an Important role in the pathogenesis of clinical developments such as in cerebral and placental malaria, in which it mediates the cytoadherence of infected red blood cells (iRBCs; infected erythrocytes) in the deep tissues. Different P/EMP1 molecules bind to various host molecules, including o2 -macroglobulin, CD36, chondroitin sulfate A (CSA), complement Iq, CR1, E-selectins and P-selectins, endothelial protein C receptor (EPCR), heparan sulfate, ICAM1, IgM, IgG, PECAM1, thrombospondin (TSP), and VCAM1. Such binding leads to activation of various host inflammatory responses. Hemoglobinopathies, including the hemoglobin C and hemoglobin S trait conditions, interfere with P/EMP1 display In knob structures of the IRBCs. This poor dsplay of P/EMP1 on the host cell surface offers protection against malaria by reducing the cytoadherence and activation of inflammatory processes that promote the development of severe disease.

[0329] Members of the large Plasmodium Interspersed repeat (pir) multigene family are named differently by parasite species, for example, yir in P. yoelii, bir in P. berghei, vir in P. vivax. Several P. falciparum gene families (stevor, rif, and P/MC-2TM) are classified with pir by their similar gene structures, which characteristically Include a short first exon, a long second exon, and a third exon encoding a transmembrane domain. In a recent study, the pir genes from P. chabaudi^df) were shown to be expressed in different cellular locations, within and on the suface of iRBCs, and in merozoites. Malaria parasites devote large portions of their genomes to gene families that ensure evasion of host immune defenses and protection of molecular processes essential to infection. These famiies emphasize the importance of research on their roles in parasite-host interactions and virulence, despite the difficulties inherent to their investigation.

[0330] An additional, exemplary polymorphic gene family comprises a group of 14 genes encoding proteins with six cysteines (6-Cys). These proteins often localize on the parasite surface interacting with host proteins and are expressed at different parasite developmental stages. 6-Cys proteins also demonstrate diverse functions and have been shown to play roles in, for example, parasite fertilization, mating interactions, evasion of immune responses, and invasion of hepatocytes. The proteins expressed in asexual stages are generally polymorphic and/or under selection, suggesting that they could be targets of the host immune response; however, their functions in parasite development remain largely unknown.

[0331] Plasmodium genomes can be highly polymorphic. Early studies demonstrated polymorphisms involving tens to hundreds of kilobases and that the chromosome structure in P. falciparum is largely conserved in central regions but extensively polymorphic is both length and sequence near the telomeres. Much of the subtelomeric variation was explained by recombination within blocks of repetitive sequences and families of genes.

[0332] The frequency of simple sequence repeats (microsatellites) in P. faldparum is estimated to be approximately one polymorphic microsatellite per kb DNA. Without wishing to be bound by any one theory, this high rate may reflect the AT -rich nature of the genome. Mlcrosatellltes seem to be less frequent In other Plasmodium species that have genomes with lower AT contents. In addition to the highly polymorphic and repetitive structure of Plasmodium genomes, there are also large numbers of Single Nucleotide Polymorphisms (SNPs) and Copy Number Variations (CNVs) (Su eta!., Plasmodium tenomks and Genetics: New Insights Into Malaria Pathogenesis, Drug Resistance, Epidemiology, and Evolution. Clin Microbiol Rev. 2019 Jul 31;32(4), which is Incorporated herein by reference In its entirety).

C. Plasmodium Proteins

1. Rh5 Invasion Complex Polypeptides Rh5 [0333] Rh5 is found In P. faldparum, but not in all other species of Plasmodium that infect humans which contain proteins that perform an analgous function as Rh5. Rh5 orthologues are also found in other species belonging to the Lavarenia subgenus, which includes parasites that infect chimpanzees and gorillas, indicating a unique role in P. falciparum invasion of human erythrocytes. See, e.g., Ragotte, et al. Trends Parasitol. 36(6) 2020, which is incorporated herein by reference in its entirety. Rh5 is expressed during the mature schizont stages and localizes to the Rhoptry, a polarized secretory organelle of P. faldparum. Rh5 is a protein that is secreted during the invasion process and is believed to be essential for the completion of parasite invasion of the erythrocyte. Rh5 functions downstream of intial parasite attachment to the erythrocyte surface and deformation of the erythrocyte membrane. Upon secretion, Rh5 binds to erythrocyte surface protein basigin, creating an attachment and a stable platform for downstream receptor-ligand interactions and associated invasion invents to occur. Bindng of Rh5 to basigin is also required for the induction of a spike in calcium within the erythrocyte, which is blocked when merozoites attempt to invade in the presence of anti-Rh5, anti-Ripr, or anti-baslgin antibodies or soluble basigin (See, e.g., Ragotte (2020), which is incorporated herein by reference in its entirety).

[0334] Rh5 can interact with multiple proteins to form complexes. The affinity of Rh5 for basigin varies based on which complex Rh5 is in. For example, Rh5 can complex with CyRPA and Ripr to form an elongated protein trimer, called RCR, on the merozoite surface that binds to erythrocyte surface protein basigin with higher affinity than Rh5 alone. (See, e.g., Ragotte (2020) and Wong (2019), each of which are incorporated herein by reference in its entirety). Rh5 has its highest affinity to basigin when interacting with PTRAMP, CSS, Ripr, and CyRPA in a pentameric complex known as PCRCR. See, Scally (2022).

[0335] Rh5 is a 63 kDa protein expressed during the mature schizont stage. It is processed and cleaved to a 45 kDa form which is shed by the parasite. The structure of Rh5 reveals a kite-like architecture formed from the coming together of two three-helical bundles. See, e.g., Ragotte (2020), which is incorporated herein by reference in its entirety.

[0336] Rh5 sequences are known (see, e.g., UnIProt accession numbers A0A159SK44, A0A159SK99, A0A159SKS8, A0A159SKW8, A0A159SL23, A0A159SL78, A0A159SL96, A0A159SLM7, A0A159SMC8, A0A159SMR9, A0A161FQT0, A0A1B1UZE2, A0A1B1UZE4, A0A1B1UZE5, A0A346RCI1, A0A346RCJ0, A0A346RCJ2, A0A346RCJ3, A0A346RCJ4, A0A346RCK4, A0A346RCK5, A0A346RCK6, A0A346RCK9, B2L3N7, Q8IFM5, each of which is incorporated herein by reference in its entirety), and exemplary Rh5 amino add sequence is provided in Table 1. Rh5-lnteractino Protein fRiorl

[0337] Rh5-lnteracting Protein (Ripr) is an approximately 120 kDa protein and localized to mkronemes during the schizont stage of the Plasmodium life cycle. The full-length 120 kDa protein is processed into two fragments of similar size, an N-termlnal fragment (Indudlng EGF domains 1 and 2) and a C-termlnal fragment (Indudlng EGF domains 3-10). Ripr colocalizes with Rh5 and CyRPA during parasite invasion at the junction between merozoites and erythrocyte. Parasites with conditional knockouts of fliRipr induce membrane deformation, but cannot complete Invasion (See, e.g., Ragotte (2020) which is incorporated herein by reference in Its entirety).

[0338] Ripr sequences are known (see, e.g., UniProt accession numbers A0A193PDI9, A0A193PDK3, A0A193PDK8, A0A193PDL3, A0A193PDL9, A0A193PDP4, A0A193PDQ8, A0A193PE01, A0A193PE05, A0A193PE07, 097302, A0A193PE17). Exemplary Ripr amino add sequences are provided In Table 1.

Cvstelne-Rich Protective Antigen fCvRPAI

[0339] Cysteine-Rich Protective Antigen (CyRPA) is a 43 kDa protein with a predicted N-terminal secretion slgial. CyRPA is a critical mediator of an Invasion multi-protein complex consisting of Rh5 and Ripr, known as RCR, that Improves Rh5 affinity to the erythrocytic basigin receptor. The ability of CyRPA to make limited contact with Ripr and Rh5 is thought to permit the disassociation of Rh5 and Ripr from this complex so they can be inserted into the erythrocyte membrane, helping to further stabilize the contact between parasite and erythrocyte. CyRPA also plays a role in the PCRCR complex which further increase Rh5 affinity to basigin. CyRPA mediated Rh5 binding to basigin is important for stabilizing the parasite-erythrocyte membrane interaction which allows downstream invasion events, like Ca2⁺ increase In the erythrocyte cytosol and establishment of tight/movlng junctions, to occur. CyRPA is highly conserved, with only a single SNP above 5% prevalence in the general population. CyRPA is important for invasion (as conditional knockdown causes the loss of invasion activity), and CyRPA has poor sero-reactivity from natural exposure (See, e.g., Ragotte (2020) and Cowman (2018) each of which is incorporated herein by reference in its entirety).

[0340] CyRPA sequences are known (see, e.g., Uniprot accession number A0A2S1Q7P0, A0A2S1Q7P5, A0A2S1Q7Q4, Q8IFM8, each of which is incorporated herein by reference in its entirety). Exemplary CyRPA amino add sequence is provided in Table 1. Ptesmtxi'u/nThrombowondic- Related Apical Merozoite Protein (PTRAMP)

[0341] Plasmodium thrombospondic-related apical merozoite protein (PTRAMP) is a 352 amino add protein that localizes to developing micronemes and relocates to the merozoite surface upon invasion and is understood to be important for host cell invasion by P. falciparum. Orthologs to the gene encoding PTRAMP is present in all malaria parasite spedes examined, indicating a conserved role in host invasion and making it an opportune target for therapeutic intervention. Important to vaccine approaches, PTRAMP appears to exhibit both linear and conformational epitopes, which would give the immune system the opportunity to utilize both B cell receptors and T cell receptors for recognition and activation when exposed to PTRAMP (Alvaro et al., Malaria Jou., 2010 Oct. 13;10.1186, which is Incorporated herein by reference in its entirety). PTRAMP can interact with cysteine-rich small secreted (CSS) protein to form a heterodimer known as PTRAMP-CSS which in turn can, through CSS, Interact with Ripr and CyRPA to form a tetrameric complex called PCRC, that ultimately Interacts with Rh5 forming a pentameric complex PCRCR which enhances Rh5 binding to host baslgin and Is important for P. falciparum to invade RBCs. Nanobodies against PTRAMP inhibit merozoite significantly reduce merozoite invasion and give evidence to the potential efficacy therapeutically targeting PTRAMP could provide (Scally et al., Nat Microb., 2022 May 4, which Is incorporated herein by reference In Its entirety).

[0342] PTRAMP sequences are known (see, e.g., Uniprot accession number [Q8I5M8]). Exemplary PTRAMP amino acid sequence is provided in Table 1.

Cvstelne-Rlch Small Secreted Protein (CSS)

[0343] Cysteine-rich small secreted protein (CSS) Is a 290 amino add cryptic 6-Cys protein comprised of two degenerate 6-Cys domains. Proteins belonging to this family commonly mediate extracellular protein-protein interactions, a role that has been confirmed to occur In P. falciparum. CSS is capable of biding both PTRAMP and Ripr. CSS Interaction with PTRAMP occurs in the endoplasmic reticulum, whereas CSS interaction with Ripr occurs after trafficking to the micronemes. CSS Interaction with PTRAMP and Ripr are involved in Invasion and the formation of the PCRC complex, which is capable of complexing with Rh5 and enhancing Rh5's Interaction with baslgin. Similar to PTRAMP, nanobodies against CSS have been showing to Inhibit Invasion, indicating Its role in the invasion process likely thought its partldpation in the PCRCR complex. Nanobodies which inhibit CSS binding to PTRAMP or Ripr do not inhibit invasion, confirming the interaction of these proteins before exposure to the merozoite surface. The Inhibitory activity of nanobodies against CSS are thought to occur by potentially blocking Ripr and/or Rh5 insertion into the RBC membrane and/or disrupting the conformational changes of the PCRCR complex required for invasion, although the exact mechanism of inhibition is not entirely known. Contrary to this observation and of immunological significance, polydonal antibodies to CSS and PTRAMP do not exhibit any neutralizing activity, indicating the immunodominance of non-neutralizing epitopes of CSS and PTRAMP that bias antibody selection (in a natural setting) away from epitopes with neutralizing capability. This observation provides the molecular rationale for identifying invasion inhibitory epitopes on CSS and PTRAMP which are highly conserved in P. falciparum, for usage in vaccine platforms. See Cowman (2022).

[0344] CSS sequences are known (see, e.g., Uniprot accession number [Q8IM47]). Exemplary CSS amino acid sequence is provided in Table 1.

[0345] In some embodiments, a Plasmodium CSS polypeptide or antigenic portion thereof comprises a serine at position 30, as numbered according to SEQ ID NO: 5. As exemplary CSS amino add sequence comprising a C30S mutation is provided in Table 1. £113

[0346] P113 is a glycosylphosphatidylinositol (GPI)-linked protein that interacts directly with the N terminus of unprocessed Rh5, which has been thought to provide a mechanism by which the Rh5 invasion complex is tethered to the merozoite surface. However, recent findings have shown that P113 may not be essential for P. falciparum growth, as other proteins and complexes exist that can perform the Rh5 membrane tethering function of P113. See, e.g., Ragotte (2020) and Scally (2022). In addition to its canonical role, P113 is present in the paristopherous vacuole (PV) and has been shown to interact with parasite export complexes like PTEX and EPIC, which are important for the transport of parasite effector proteins across the paristopherous vacuole membrane (PVM). P113 has also been shown to play a role in PVM architecture, as the deletion of this gene leads to a disruption In PVM morphology. P113 orthologues are found in all Plasmodium species sequenced thus far, which is suggestive of common and conserved functions (Bullen et al. (2022) Molecular Microbiology 117:1245-1262, which is incorporated herein by reference in its entirety). Despite this fact, in P. berghei, Pl 13 knockout parasites were viable, indicating the protein was not essential for asexual blood stage growth and invasion. The knockout parasites do, however, display defects In natural sporozoite transmission, leading to delayed patency in Infected mice (Offeddu et al. (2014) Mol. Blochem. Parasitology 193: 101-109, which is incorporated herein by reference in its entirety).

[0347] Plasmodium Pl 13 sequences are known (see, e.g., Uniprot accession number Q8ILP3). Exemplary P113 amino acid sequence is provided in Table 1.

Table 1: Exemplary Sequences for Rh5 Invasion Complex Polypeptides

SEQ ID Protein Sequence (Amino Add) NO:

1 Rh5 (3D7) MIRIKKKLILTIIYIHLFILNRLSFENAIKKTKNQENNLTLLPIKSTEEEKDDIKNGKDIKKEI DNDKENIKTNNAKDHSTYIKSYLNTNVNDGLKYLFIPSHNSFIKKYSVFNQINDGMLLNE KNDVKNNEDYKNVDYKNVNFLQYHFKELSNYNIANSIDILQEKEGHLDFVIIPHYTFLDYY KHLSYNSIYHKSSTYGKCIAVDAFIKKINETYDKVKSKCNDIKNDLIATIKKLEHPYDINNK NDDSYRYDISEEIDDKSEETDDETEEVEDSIQDTDSNHTPSNKKKNDLMNRTFKKMMDE YNTKKKKLIKCIKNHENDFNKICMDMKNYGTNLFEQLSCYNNNFCNTNGIRYHYDEYIHK ULSVKSKNLNKDLSDMTNILQQSELLLTNLNKKMGSYIYIDTIKFIHKEMKHIFNRIEYHT KIINDKTKIIQDKIKLNIWRTFQKDELLKRILDMSNEYSLF1TSDHLRQMLYNTFYSKEKHL NNIFHHUYVLQMKFNDVPIKMEYFQTYKKNKPLTQ

2 Ripr (3D7) MFRIFFTLLIIIUKKTSAIDLIEGIFYEKNEIDKLTFSLDHRVRDNLKTDLILNNNGENDYAY LNKYVYniNRDSTEKIKTFFSHNKDMKSCDYFISKEYNSSDKTNQICYKKTFCGWIPNS EBKTNKITNDKLYCAHFNSTHIIIYYISQPLLLEPHVVYEETFFEKGKNDQINCQGMYISL RSVHVHTHNAILQQETLTfTKNLCDGKNNCKFDFDSIKYENKSLTHYLFFINIQYQCISPL NLQENEMCDVYNDDTHKATCKYGFNKIELLKNVCEENYRCTQDICSVNQFCDGENETCT CKTSLLPSAKNNCEYNDLCTVLNCPENSTCEQIGNGKKAECKCENGKYYHNNKCYTKND LELAIKIEPHKKEKFYKNNLYQGKALKPEYIFMQCENGFSIEVINAYVSCYRVSFNLNKLKY VTESLKKMCDGKTKCAYGNTIDPIDDLNHHNICNNFNTIFKYDYLCVFNNQNITSDKNSH LHSNIPSLYNSSILPDINKSKFHLISRNSRTNQYPHNNISMLEIQNEISSHNSNQFSTDPH TNSNNINNMNIKKVEIFRSRFSSKLQCQGGKINIDKAILKGGEGCNDLLLTNSLKSYCNDL SECDIGUYHFDTYCINDQYLFVSYSCSNLCNKCHNNSTCYGNRFNYDCFCDNPYISKYG NKLCERPNDCESVLCSQNQVCQILPNDKLICQCEEGYKNVKGKCVPDNKCDLSCPSNKV CVIENGKQTCKCSERFVLENGVCICANDYKMEDGINCIAKNKCKRKEYENICTNPNEMCA YNEETDIVKCECKEHYYRSSRGECILNDYCKDINCKENEECSIVNFKPECVCKENLKKNNK GECIYENSCLINEGNCPKDSKCIYREYKPHECVCNKQGHVAVNGKCVLEDKCVHNKKCSE NSICVNVMNKEPICVCTYNYYKKDGVCLIQNPCLKDNGGCSRNSECTFKYSKINCTCKEN YKNKDDSCVPNTNEYDESFTFQYNDDASIILGACGMIEFSYIYNQIIWKINNSKESYVFYY DYPTAGNIEVQIKNEIFHTHYLKKKIGNSVIYDDFQVDHQTCIYENVFYYSNQN

3 CyRPA (3D7) MIIPFHKKFISFFQIVLWLLLCRSINCDSRHVFIRTELSFIKNNVPCIRDMFFIYKRELYNIC

LDDLKGEEDETHIYVQKKVKDSWrTLNDLFKETDLTGRPHIFAYVDVEEIIILLCEDEEFSN

0. Additional Plasmodium Proteins

[0348] Plasmodium parasites are known to express various proteins at different stages of their lifecycles. Exemplary malarial proteins are described below, and exemplary amino add sequences are provided in Table 2. Circumsporozoite Protein fCSPI

[0348] Circumsporozoite protein (CSP) is a multifunctional protein that is involved in Plasmodium life cycle, as it is required for the formation of sporozoites in the mosquito midgut, the release of sporozoites from the oocyst, invasion of salivary glands, attachment of sporozoites to hepatocytes in the liver, and sporozoite invasion of hepatocytes (see, e.g., Zhao et al. (2016) PLoS ONE 11(8): e0161607, which is incorporated herein by reference in its entirety). CSP is present in all Plasmodium spedes, and although variation exists in the amino add sequence across species, the overall domain structure of a central repeat region and nonrepeat flanking regions is well conserved (see, e.g., Zhao et al. (2016) PLoS ONE 11(8): e0161607; Wahl et al. (2022) J. Exp. Med. 219: e20201313, each of which is incorporated herein by reference in its entirety). CSP sequences are known (see, e.g., UniProt accession numbers A0A2L1CF52, A0A2L,lCF88, C6FGZ3, C6FH2,7 C6FHG7, M1V060, M1V0A3, Ml V0 B0, M1V0C4, M1V0E0, M1V9I4, M1VFN9, M1VKZ2, P02893, Q5EU9, Q5BK2, Q5BK8, Q5EIL3, Q5EIL5, Q5BL8, Q5R2L2, Q7K740, Q8I9G5, Q8I9J3, Q8I9J4). Exemplary CSP amino acid sequences are provided in Table 2. E140 [0350] E140 is found in every Plasmodium species for which a genomic sequence is available, and is well conserved, with amino add identity ranging from 34-92% among spedes. See, e.g., Smith , et al. PLoS one 15.5 (2020): 60232234; http://doi: 10.1371/joumal.pone.023223; and U.S. Patent Publication No. US 2019/0117752; each of which are incorporated herein by reference in their entirety. E140 is also highly conserved (95-99%) in P. falciparum sir aixxs isolated from different locations around the world, and exhibits a low mutation frequency. E140 is expressed at different life stages of malaria parasites (spedfically, E140 has been detected in sporozoites, liver, and blood stage parasites).

[0351] Protein structure algorithms predict that the E140 protein has five transmembrane domains, presumable spanring a parasite or host-derived membrane. E140 displays distinct patterns of protein expression In mature sporozoites, late liver, and late schizont stages. It traffics to the anterior and posterior ends of the sporozoite, the parasitophorous vacuole space of the late liver stage and around developing merozoites in the late schizont stage. It is also known to be expressed In mature salivary gland sporozoites as well as oocyst-derived sporozoites and oocysts.

[0352] E140 sequences are known (see, e.g., UnIProt accession numbers A0A650D649, A0A650D653,

A0A650D672, A0A650D687, AOA650D690, A0A650D694, A0A650D6A3, A0A650D6B8, A0A650D6L3, A0A650D6L7, Q8I299, each of which Is incorporated herein by reference In its entirety), and an exemplary E140 amino add sequence Is provided In Table 2.

CeiTOS

[0353] CeiTOS Is required for sporozoite traversal through Kupffer cells during the liver invasion process.

CeiTOS forms a pore from within the cell, allowing for sporozoite egress into the liver. Antibody epitopes have been characterized from Immunized mice and infected human populations (PFand PV). In mouse studies, Immunization with CeiTOS has been shown to provide protection and against challenge. Vaccination with CeiTOS may generate antibodies that can bind the extracellular domain of the pore-forming complex, blocking complete formation of the pore and preventing sporozoite traversal Into the liver. See, e.g., Jimah et al., ELIFEe2016 Dec l;5:e20621. dol: 10.7554/eUfe.20621, which is incorporated herein by reference in its entirety.

[0354] CeiTOS sequences are known (see, e.g., Uniprot accession number M1ETJ8, Q53UB7, A0A2R4QLA5, A0A2R4QU0, A0A2R4QLI5, A0A2R4QU1, A0A2R4QU4, M1ETJ8, Q53UB8, Q8I5P1, each of which is incorporated herein by reference in its entirety). An exemplary CeiTOS amino add sequence is provided in Table 2.

[0355] SPECT1 and SPECT2 (the latter also sometimes referred to as perforin-like protein 1 (PLP1)) are siyiificant Plasmodium proteins that may play a role in cell traversal. See Yang et al., Cell Rep. 2017 Mar 28;18(13):3105-3116. doi: 10.1016/j.celrep.2017.03.017, which is incorporated herein by reference in its entirety. Targeted disruption of P. falciparum SPECT1 or SPECT2 has been shown to reduce infectivity of sporozoites in liverstage development in humanized mice. However, mechanisms of cell traversal of these two proteins are yet to be defined in P. falciparum. See Yang et al.

[0356] SPECT1 and SPECT2 are considered attractive pre-erythrocytic immune targets due to the key role they are thought to play in the crossing of the malaria parasite across the dermis and the liver sinusoidal wall, prior to invasion of hepatocytes. Recombinant P. falciparum SPECT2 has been shown to cause lysis of red blood cells in a Ca^2-* -dependent manner, as has the MACPF/CDC domain of ASPECT2. ASPECT2 has also been implicated in the Ca2+ -dependent egress of P. falciparum merozoites from red blood cels.

[0357] SPECT1 and SPECT? sequences are known (see, e.g., UniProt accession numbers Q8IDR4 and Q9U0J9, each of which is incorporated herein by reference in its entirety), and exemplary amino add sequences for SPECT1 and SPEC!? are provided in Table 2.

Exported Protein 1 (EXP 1

[0358] Exported protein 1 (EXP1) is a single pass transmembrane protein with an N-terminal signal peptide expressed during intraerythrocytlc stage and liver stage (see, e.g., Spielmann et al., Int J Med Microbiol. 2012 Oct;302(4-5): 179-86, which is Incorporated herein by reference In its entirety). EXP1 was shown to Initially localize to dense granules in merozoites and then be transported to parasltophorous vacuolar membrane (PVM) after invasion (see, e.g., Iriko et al., Parasitol Int. 2018 Oct;67(5):637-639, which is incorporated herein by reference in its entirety). Once localized to the PVM, EXP1 forms homo-oligomers with a N-terminus that is exposed to the parasltophorous vacuolar lumen and a C-terminus that is exposed to the red blood cell cytosol (see, e.g., Mesén- Ramrez et al., PLoS Biol. 2019 Sep 30;17(9):e3000473, which is Incorporated herein by reference in Its entirety).

[0359] EXP1 has been demonstrated to possess glutathione S-transferase (GST) activity that may protect Plasmodium from oxidative damage (see, e.g., Mesén-Ramirez et al., PLoS Biol 17(9) 2019 Sep 30;17(9):e3000473, which is incorporated herein by reference in its entirety). Recently, it was demonstrated that EXP1 is important for Plasmodium survival by maintaining correct localization of EXP2, a nutrient-permeable channel in the PVM (see, e.g., Mesen-Rami'rez et al., PLoS Biol. 2019 Sep 30;17(9):e3000473, which is Incorporated herein by reference in Its entirety).

[0360] P. falciparum EXP1 polypeptide sequences are known (see, e.g., UniProt accession number Q8IIF0,

W7JTD3, Q25840, Q548U2, Q5VKK2, Q5VKK5, Q5WRH8, Q6V9G4, Q6V9G6, Q6V9G9, Q6V9H1, Q6V9H2, Q9U590, P04923, P04926, each of which is Incorporated herein by reference in its entirety). An exemplary EXP1 amino add sequence Is provided in Table 2.

Unregulated In Infective Sporozoites Gene 3 (UIS3)

[0361] Upregulated in infective sporozoites gene 3 (UIS3) is a membrane-bouxi protein localized to sporozoite parasltophorous vacuolar membrane (PVM) in infected hepatocytes. UIS3 was shown to interact with liver fatty addbinding protein (L-FABP) and be involved in fatty acid and/or lipid import during phases of Plasmodium growth (see, e.g., Sharma et al., J Biol Chem. 2008 Aug 29; 283(35): 24077-24088; Mikolajczak et al., Int J Parasitol. 2007 Apr;37(5):483-9, each of which is incorporated herein by reference in its entirety).

[0362] After sporozoite invasion of host liver cells, there Is synthesis of vital Plasmodium structural features (e.g., parasltophorous vacuolar membrane). During hepatocytic stages, the Plasmodium relies on host fatty adds for rapid synthesis of its membranes (see, e.g., Sharma et al., J Biol Chem. 2008 Aug 29; 283(35): 24077-24088, which is incorporated herein by reference in its entirety). UIS3 insertion in the PVM provides Plasmodium a method to import essential fatty adds and/or lipids during rapid sporozoites growth phases (see, e.g., Sharma et al., J Biol Chem. 2008 Aug 29; 283(35): 24077-24088, which is incorporated herein by reference in its entirety).

[0363] Immunization with UIS3-defident Piasmodum berghei sporozoites protected against malaria in rodent malaria model (see, e.g., Mueller et al., Nature. 2005 Jan 13;433(7022): 164-7, which is incorporated herein by reference in its entirety). UIS3-defident Plasmodium berghei can start the transformation process in the liver; however, they show severe defects during transformation into trophozoites (see, e.g., Mueller et al., Nature. 2005 Jan 13;433(7022): 164-7, which is incorporated herein by reference in its entirety). UIS3-deficient Plasmodium berghei are also unable to develop into mature liver schizonts and therefore abort malaria infection within the liver itself (see, e.g., Mueller et al., Nature. 2005 Jan 13;433(7022): 164-7, which is incorporated herein by reference in its entirety). Further, it was previously demonstrated that UIS3 derived from Plasmodium berghei and UIS3 derived from Plasmodium faldparum exhibited a low (i.e. 34%) amino acid sequence identity (see, e.g., Mueller et al., Nature. 2005 Jan 13;433(7022): 164-7, which is incorporated herein by reference in its entirety).

[0364] Plasmodium UIS3 sequences are known (see, e.g., UniProt accession number A0A509ARS3, A0A1C6YLP3, Q8IEU1, A0A384KU1, A0A1G4H423, A0A077YB01, Q9NFU4, each of which is Incorporated herein by reference In Its entirety). An exemplary U1S3 amino acid sequence Is provided In Table 2.

Early Transcribed Membrane Protein 10.3 (ETRAMP10.3) and Unregulated in Infective S es Gene 4 (UIS4)

[0365] Upregulated in infective sporozoites gene 4 (UIS4) contains a single transmembrane domain and localizes to secretory organelles of sporozoites and to the parasitophorous vacuole membrane (PVM) of liver stages. UIS4 is not expressed In blood stages or early sporozoites that are produced in oocysts (see, e.g., Mackellar et al., Eukaryot Cell. 2010 May; 9(5): 784-794, which is incorporated herein by reference in Its entirety).

[0366] Deletion of UIS4 gene is associated with arrest of early liver stage development (see, e.g., Vaughan and Kappe, Cold Spring Harb Perspect Med. 2017 Jun l;7(6):a025486, which Is Incorporated herein by reference in its entirety). Recently, UIS4 was demonstrated to be Involved in Plasmodium berghei survival by eluding host actin structures deployed as part of host cytosolic defense (see, e.g., Sana et al., (Science. 2022 Apr 22;25(5): 104281. doi: 10.1016/j.isci.2022.104281. eCollection 2022 May 20, which Is Incorporated herein by reference In Its entirety). P. falciparum has an ortholog to UIS4 named ETRAMP10.3 which is not able serve as a functional compliment to P. yoeliiUIS4, Indicating It likely serves a different function In P. falciparum's lfie cycle (see Mackdlar et al., Eukaryot Cell 9:784-94 (2010), which is incorporated herein by reference In its entirety).

[0367] Plasmodium faldparum early transcribed membrane protein 10.3 (ETRAMP10.3) Is an approximately 10 kDa protein and member of the early transcribed membrane proteins multigene family, a family which is conserved across Plasmodium species and includes proteins located In the parasitophorous vacuole. Several ETRAMP proteins are specific to P. faldparum and not found In Plasmodium species that infect other organisms. ETRAMP10.3 is one example, which is expressed in both liver and blood stage P. faldparum parasites. ETRAMP10.3 transcription has been found to peak during the transition from ring to trophozoite stages of P. faldparum blood stage infection in a human host ETRAMP10.3 localizes to the parasitophorous vacuole and is exported to a host erythrocyte during blood stage infection. Although ETRAMP10.3 is sometimes referred to as Upregulated in Infectious Sporozoites gene 4 (UIS4), ETRAMP10.3 is understood to be an ortholog of UIS4 on the basis of synteny and structural similarity.

However, ETRAMP10.3 is not a functional ortholog of UIS4 and may play a different biological role. Although the biological function of ETRAMP10.3 has not yet been completely resolved, localization to vesicular structures in the host erythrocyte suggests a role in host-parasite interaction or in remodeling of infected erythrocyte. ETRAMP10.3 appears to play a key role in the Plasmodium life cycle. When ETRAMP10.3 is deleted, the deletion can lead to the disruption of liver-stage development in mice and asexual blood stage progression.

[0368] Although the terms “UIS4" and "ETRAMP10.3” are in the literature sometimes used to refer to different proteins, in context of the present disclosure, the terms "UIS4* and "ETRAMP10.3" interchangeably to refer to ETRAMP10.3. [0369] Plasmodium UIS4 sequences are known (see, e.g., UniProt accession number Q8DM9, which is incorporated herein by reference in its entirety). An exemplary UIS4 amino add sequence is provided in Table 2. Liver Specific Protein 1 fUSP-1)

[0370] Liver specific protein 1 (LISP-1) is expressed during Plasmodium development in hepatocytes and localized to the parasitophorous vacuolar membrane (PVM) (see, e.g., Ishino et al., Cell Microbiol. 2009 Sep; 11(9): 1329-1339, which is incorporated herein by reference in its entirety). LISP-1 was shown to be expressed at high levels during late liver stages development and to be involved in PVM breakdown and subsequent merozoite release (see, e.g., Ishino et al., Cell Microbiol. 2009 Sep; 11(9): 1329-1339, which is incorporated herein by reference in its entirety).

[0371] Intracellular Plasmodium deficient In LISP-1 develop into hepatic merozoites and display normal infectivity to erythrocytes (see, e.g., Ishino et al., Cell Microbiol. 2009 Sep; 11(9): 1329-1339, which is incorporated herein by reference in its entirety). However, USPl-defident liver-stage Plasmodium do not rupture PVM and remain trapped inside hepatocytes (see, e.g., Ishino et al., Cell Microbiol. 2009 Sep; 11(9): 1329-1339, which is incorporated hereto by reference in its entirety).

[0372] Plasmodium LISP-1 sequences are known (see, e.g., UniProt accession number A0A210C2X6, Q8ILR5, each of which is incorporated herein by reference in its entirety). An exemplary LISP-1 amino add sequence is provided In Table 2.

Liver Specific Protein 2 fLISP-2)

[0373] Liver spedfic protein 2 (LISP-2) contains a modified 6-cys domain and is expressed during Plasmodium development in hepatocytes (see, e.g., Orito et al., Mol Microbiol. 2013 Jan;87(l):66-79, which is incorporated herein by reference In Its entirety). LISP-2 was shown to be expressed by liver stages Plasmodium, exported to hepatocytes, and be distributed throughout the host cell, including the nucleus (see, e.g., Orito et al., Mol Microbiol. 2013 Jan;87(l): 66-79, which is incorporated herein by reference In Its entirety).

[0374] Intracellular Plasmodium deficient in LISP2 do not mature effectively during merozoites development (see, e.g., Orito et al., Mol Microbiol. 2013 Jan;87(l):66-79, which is incorporated herein by reference In its entirety).

[0375] Plasmodium LISP-2 sequences are known (see, e.g., UniProt accession number A0A2I0BZR4, Q8I1X6, Q9U0D4, each of which is Incorporated hereto by reference in its entirety). An exemplary LISP-2 amino add sequence is provided in Table 2.

Thrombosoondin-Related Adhesion Protein fTRAP)

[0376] Thrombospondin-related adhesion protein (TRAP) contains an N-terminal domain that is commonly referred to as von Willebrand factor A domain, although it is most similar to an integrin I domain because it contains a metal Ion-dependent adhesion site (MIDAS) with a bound Mg^2t Ion that Is required for sporozoite motility In vitro and infection in vivo (see, e.g., Lu et al., PLoS One. 2020; 15(1): e0216260, which is incorporated herein by reference in its entirety). The I domain is inserted in an extensible p-ribbon and followed by a thrombospondin repeat (TSR) domain, a proline-rich segment at the C-terminus, a single-pass transmembrane domain, and a cytoplasmic domain (see, e.g., Lu et al., PLoS One. 2020; 15(1): e0216260, which is incorporated herein by reference in its entirety). Sequence analysis of the proline-rich segment revealed the presence of SH3-domain binding PxxP motifs in PlasmodiumTRAPs (Akhouri et al., Malar J. 2008 Apr 22;7:63. doi: 10.1186/1475-2875-7-63, which is incorporated herein by reference in its entirety). [0377] TRAP is stored in the mkronemes and becomes surface exposed at the sporozoite anterior tip when parasite comes in contact with host cells (Akhouri et al., Malar J. 2008 Apr 22;7:63. doi: 10.1186/1475-2875-7-63, which is incorporated herein by reference in its entirety). TRAP also plays an important role in liver cell invasion of sporozoites by helping sporozoites in gliding motility and in recognition of host receptors on the mosquito salivary gland and hepatocytes (Akhouri et al., Malar J. 2008 Apr 22;7:63. doi: 10.1186/1475-2875-7-63, which is incorporated hereto by reference in its entirety).

[0378] PlasmodiumTRAP sequences are known (see, e.g., UniProt accession numbers A0A5Q2EXK8, A0A5Q2EZD7, A0A5Q2F1F6, A0A5Q2F2B8, A0A5Q2F2H6, A0A5Q2F4G9, 076110, P16893, Q01507, Q26020, Q76NM2, W8VNB6, each of which is Incorporated herein by reference in its entirety), and an exemplary TRAP amino add sequence Is provided In Table 2.

Liver-Staoe-Assodated Protein-1 (LSAP-1)

[0379] Liver-stage-associated protein (LSAP-1) has been shown to be found mainly at the periphery of the Intracellular hepatic parasite throughout its development, but not in blood stage parasites and possibly in minor quantities In salivary gland sporozoites (see, e.g., Siau et al., PLoS Pathog. 2008 Aug 8;4(8):el000121, which Is Incorporated herein by reference In Its entirety). LSAP-1 is among the most abundant transcripts In the salivary gland transcriptome but has not been detected in proteomlc surveys of sporozoites. Rather, expression has only been detected only In liver stages (see, e.g., Slau et al., PLoS Pathog. 2008 Aug 8;4(8):el000121, which is Incorporated herein by reference In its entirety).

[0380] Plasmodium LSAP-1 sequences are known (see, e.g., UniProt accession number Q8I632, W7JR53, each of which Is incorporated herein by reference in Its entirety). An exemplary LSAP-1 amino add sequence is provided In Table 2.

Liver-Stage-! "Delated Protein-2 fLSAP-2)

[0381] Like LSAP-1, LSAP-2 Is also among the most abundant transcripts In the salivary gland transcriptome but has not been detected in proteomlc surveys of sporozoites. LSAP-2 has shown some efficacy as a vaccine when combined with other antigens. See, e.g., Halbroth et al., Infect Immun. 2020 Jan 22;88(2):e00573-19. doi: 10.1128/IAI.00573-19. Print 2020 Jan 22, which Is incorporated herein by reference In its entirety.

[0382] Plasmodium LSAP-2 sequences are known (see, e.g., UniProt accession number Q8I632, W7JR53, each of which is incorporated herein by reference in its entirety). An exemplary LSAP-2 amino add sequence is provided in Table 2.

Liver-Staae Antigen 1 (LSA-1)

[0383] Liver-Stage Antigen 1 (LSA-1) is expressed after Plasmodium have invaded hepatocytes and antigen accumulates in the parasitophorous vacuole (see, e.g., Tucker, K. et al., 2016, 'Pre-Erythrocytic Vaodne Candidates in Malaria', in A. J. Rodriguez-Morales (ed.), Current Topics in Malaria, IntechOpen, London. 10.5772/65592, each of which is incorporated herein by reference in its entirety). The function of LSA-1 remains currently not known (see, e.g., Tucker, K. et al., 2016, 'Pre-Erythrocytic Vacdne Candidates in Malaria', in A. J. Rodriguez-Morales (ed.), Cun-ent Topics in Malaria, IntechOpen, London. 10.5772/65592, which is incorporated herein by reference in its entirety).

[0384] LSA-1 is a 230 kDa preerythrocytic stage protein containing a large central region consisting of over eighty 17 amino add residue repeat units flanked by highly conserved C- and N-termlnal regions (Richie, T.L. and Parekh, F.K. (2009) Malaria, which is incorporated herein by reference in its entirety). In Vaccines for Biodefense and Emerging and Neglected Diseases (Barrett, A.D.T. and Stanberry L.R., eds), pp. 1309-1364, Elsevier, which is incorporated herein by reference in its entirety). LSA1 is expressed only by liver stage Plasmodium and not by sporozoites (Richie, T.L. and Parekh, F.K. (2009) Malaria, which is incorporated herein by reference in its entirety). In Vaccines for Biodefense and Emerging and Neglected Diseases (Barrett, A.D.T. and Stanberry L.R., eds), pp. 1309- 1364, Elsevier, which is incorporated herein by reference in its entirety). The repeat region results in significant variation of the protein between strains of Plasmodium falciparum (see, e.g., Tucker, K. et al., 2016, 'Pre-Erythrocytic Vaccine Candidates in Malaria', in A. J. Rodriguez-Morales (ed.), Current Topics in Malaria, IntechOpen, London. 10.5772/65592, which is incorporated herein by reference in its entirety).

[0385] Plasmodium LSA-1 sequences are known (see, e.g., UniProt accession number Q25886, Q25887, Q25893, Q26028, Q9GTX5, 096125, each of which Is Incorporated herein by reference In Its entirety). An exemplary LSA-1 amino acid sequence is provided in Table 2.

Liver stage antigen 3 (LSA-3)

[0386] Liver stage antigen 3 (LSA-3) Is a 200-kDa protein that Is composed of three nonrepeating regions (NR- A, NR-B, and NR-C) flanking two short repeat regions and one long repeat region (see, e.g., Tucker, K. et al., 2016, 'Pre-Erythrocytic Vaccine Candidates in Malaria', in A. J. Rodriguez-Morales (ed.), which Is incorporated herein by reference in its entirety), Current Topics in Malaria, IntechOpen, London. 10.5772/65592, which is incorporated herein by reference in its entirety). The nonrepeat regions are well conserved across geographically diverse strains of Plasmodium faidparum (see, e.g., Tucker, K. et al., 2016, 'Pre-Erythrocytic Vaccine Candidates in Malaria', in A. J. Rodriguez-Morales (ed.), Current Topics In Malaria, IntechOpen, London. 10.5772/65592, which is Incorporated herein by reference in Its entirety). The most significant variation Is In the repeating regions due to organization and number of repeating subunits rather than composition of the repeating regions (see, e.g., Tucker, K. et al., 2016, 'Pre-Erythrocytic Vaccine Candidates In Malaria', in A. J. Rodriguez-Morales (ed.), Current Topics in Malaria, IntechOpen, London. 10.5772/65592, which is Incorporated herein by reference In its entirety).

[0387] Recently, in vitro data has shown that antibodies against LSA-3 (in particular, the C-termlnal portion of LSA-3) may provide some protection (see, e.g., Morita et al, Sci Rep. 2017 Apr 5;7:46086. doi: 10.1038/srep46086, which is incorporated herein by reference in Its entirety).

[0388] Plasmodium LSA-3 sequences are known (see, e.g., UniProt accession number C7DU21, C7DU22, C7DU23, C7DU24, C7DU25, C7DU26, C7DU27, C7DU28, C7DU29, C7DU32, C7DU33, C7DU34, C7DU36, C7DU37, C7DU38, C7DU39, C7DU40, Q8I042, Q8I0A5, Q8I0D0, Q8IFR1, Q8IFR2, Q8IFR3, Q8IFR4, Q8IFR5, Q8IFR6, Q8IFR7, Q8IFR8, Q8IFR9, Q8IFS0, Q8IFS1, Q8IFS2, Q8IFS3, Q8IFS4, Q8IFS5, Q8IFS6, Q8IFS7, Q8IFS8, Q8IFS9, Q8IFT0, Q8IFT1, Q8IFT2, Q8IFT3, Q8IFT4, Q9U0N9, Q9U0P0, A0A2I0BVD6, A0PFM9, 096275, each of which is incorporated herein by reference In its entirety). An exemplary LSA-3 amino acid sequence is provided In Table 2.

Glutamic Add-Rich Protein

[0389] Glutamic add-rich protein (GARP) is a 80kDA protein which derives its name from its glutamic rich amino add sequence which comprises 24% of all its residues. GARP Is predominantly expressed in ring stages and trophozoites and has been shown to be a non-essential gene in cell culture but highly immunogenic in animal models. Although GARP is non-essential in cell culture, its localization to the periphery of infected erythrocytes may indicate a role in the sequestration of infected erythrocytes. CARP'S involvement in sequestration has been proposed to occur by way of binding with an chloride/bicarbonate anion exchanger. Antibodies against GARP have been proposed to serve as signatures of protection against severe malaria and have shown efficacy in experimental trials

SEQ ID Protein Sequence (Amino Add) NO:

7 CSP MMRKLAILSVSSFLFVEALFQEYQCYGSSSNTRVLNELNYDNAGTNLYNELEMNYYGKQE NWYSLKKNSRSLGENDDGNNEDNEKLRKPKHKKLKQPADGNPDPNANPNVDPNANPN VDPNANPNVDPNANPNANPNANPNANPNANPNANPNANPNANPNANPNANPNANPNA NPNANPNANPNANPNANPNANPNVDPNANPNANPNANPNANPNANPNANPNANPNAN PNANPNANPNANPNANPNANPNANPNANPNANPNANPNANPNKNNQGNGQGHNMPN DPNRNVDENANANSAVKNNNNEEPSDKH1KEYLNKIQNSLSTEWSPCSVTCGNGIQVRI KPGSANKPKDELDYANDIEKKICKMEKCSSVFNWNSSIGLIMVLSFLFLN

8 TRAP MNHLGNVKYLVIVFUFFDLFLVNGRDVQNNIVDEIKYREEVCNDEVDLYLLMDCSGSIRR HNWVNHAVPLAMKUQQLNLNDNAIHLYASVFSNNAREIIRLHSDASKNKEKAUIIKSLL STNLPYGKTNLTDALLQVRKHLNDRINRENANQLWILTDGIPDSIQDSLKESRKLSDRG VKIAVFGIGQGINVAFNRFLVGCHPSDGKCNLYADSAWENVKNVIGPFMKAVCVEVEKTA SCGVWDEWSPCSVTCGKGTRSRKREILHEGCTSELQEQCEEERCLPKREPLDVPDEPED DQPRPRGDNFAVEKPNENIIDNNPQEPSPNPEEGKGENPNGFDLDENPENPPNPPNPPN PPNPPNPPNPDIPEQEPNIPEDSEKEVPSDVPKNPEDDREENFDIPKKPENKHDNQNNLP NDKSDRYIPYSPLSPKVLDNERKQSDPQSQDNNGNRHVPNSEDRETRPHGRNNENRSY PVlDLSQNNFQVNlYMHSKFlPNDYSLRKDIVPHmNNVPSNVFILSLKELLKTIFIQHME KSQTTTFQLNLYDFEVEFFILTFNRFTSKFHKMNSNVLSNLTISNFYSTYIHGGLFLNIDK1 KIDIPNLLNDFIDHFSKWIDFSHHNIVKQ _

16 USP2 (3D7) MEKGSILSFIFFCSWIFIRFIGYFFCNRYMTEEPYNNIFBIKPENLYSSLLLLSNEKENDFP SSTNCNGYMKCIPFYNNVSERWKRYNFIQLYIIRSALNIHVMSKYNMLNKYNKETMRLLK RNNNVENRINNISNHYLCSGFKKENRLFFI±FYIG1KMMKLYIRNLFMKYIKIYYKTKHFEK NIETMKKWYVERDNLFDIERNNLFDILYMLKRIDSYVKNIYSIISNNFLYVIRIIFLPFEKIY FSLKSUMIKKMNMSSSYYYYYVNMFSLYKKNYNKYEEIFIHEQRVIYPNEYLKNEMLDKY RRVIRILSGQHDNPFIDSLLINPEKIEKDDLDVKQKKKKIIEELKKKKENTNTNTNTSTNTS ANTNTSTNTSANTNTSTKESHILDESKLETFYRDELDKMGKEBETYFKGNIDKKSLDEFH KILLEELNKMDKDELYEMYREELNRIEQEKIRNMNKEEINKTYKDBNNMNSDQVDKIHR EELEK1EKEKINKMDKDEIDKIYREELDKMDRDAIYSMYIEDISNKN1KDL1KNEKETNKDK NKKKDIDINKKKKKDIDIDVDIDKDIHKDHVEELYGEVKNKLSKEELDRMDRDALYRVYL EELDRMNRDELYRVYLEELEKIDKEEKEKIHREKLHKIEKEKINKMDKDQIDKIYEEELNK MDSDEIQHVRRAILQDIQKEKIQNLELEBDRLYKEELDRMDREARYEIPMRNLSRNEKD NIIHRNIKNESNQKNKKENVNVFIIHDNNDSNNNNNNNNRDVNNLNNKHTNNNYNENV EVELWRNLDKDKGAKIEDIIDYFNKEIKKDKNVNVSNIVNFLNSKVGKDNTPIQHKKEN QVDWRKNIQIIQEDNIKNKGQKDNTEMLDNNKEITNID1KNVDDIKNVGDIKSVGDIKS VDDINNVDGIKNVDGIKNVDGIKNVDGINNVGDINNAGDTNNAGDINNVGDINNSVDIY NVEHIDEAEKKPNLDNPKKFDWTQVFKDKVTEKIKNEEKFNNSKENIQNDIRDKEIHKD DRIKGITSREKNAEEINNNEKKDKFVYEFYTSNKKENIDKEEENNIDDKNIKIEIEPNYEIN NNFEEENKNEINVIIDKEAKNNMDKDDSNNNNNIQKNNIIIKDNTNVSEEVHITESSKEIA EFFNNIIKNSNILDMCSKMNASDSEKGFICINGNNYIINPGTYHIINIKYPDYNNVRKKWY DSMDCISINNKDENNNNKEHNYYNKNDDDLYLKKSVEEFIPGFLSNINKVDDLARIFTPS FIQNDIFLNCIYKYRNDFDKNNNIYSFPMK1FLRKNSTKIKGCSFQIDEDPLLYKDYSEKES FLSNKIILNNSNRNTECVLHASNEIVGFQCGPPYKSYDNIQYRHLTNKSNDIQKNIFGIYS NNNSSYSHLFKNIFNNEHKLYNVGGYFQTEPINCFEFVNDNINVEDILPGAVPFPRFDLIH HDLDVNQTRYILLNETNQDKTISCTCNYFTEPNIVYTGKIIIKVEEEKIYICrKKLQTEFNDI INNKKEIYHEKKMNHIIKEKKDEDENDMSFNKNYVNNYNENFKINDINNFYNRNNHPNN NYHNDYHNNHSSKGNHTNKIHDTFLKNKYNISFNNLKFYNIKHENKNNQDVINYEYNVD DYHEVQDGQDESFKEEEDFJDFKENIINDNTNHNNTDLDDDKYNKYHNNNNNNNNNSS FKSIESNLDLQRSILQSGDTQQVWINKSKNVNVLFPDKKKNHTHPNEEKRTFPKYISLVY KEKNKDNEKIDICTLTHKEFYPPLLRGGKDSDPENKQTGVEINNGVEKKNDVQIKNDVEI KNDVEIKNDVBNDDVEIKNDVE1KNDVBKNDVEINDDVBNDDVEINNGVEINDGVEN KDN1HEGNNNLENDSFNEDT1EEPFENIFDFINEETSSNENSBILDSADSIKRKLGHNFLD IISAGKFKIRHKEKKTKNKKKK _

17 EXP1 (3D7) MKJLSVFFLALFHIFNKESLAEKTNKGTCSGVSSKKKNKKGSGEPUDVHDUSDMIKKEE ELVEVNKRKSKYKLATSVLAGLLGWSTVLLGGVGLVLYNTEKGRHPFKIGSSDPADNANP DADSESNGEPNAGPQVTAQDVTPEQPQGDDNNLVSGTEH _

18 EXP2 (3D7) MKVSYIFSFFLLFFVYKNTNTWCDNGYGDLAATSALTTVIKDPISLTIKDIYEHGVKNPFT KIIHKLKKHRYRKVLRWSRMWWVLLVREIVGDNnEKKTEKALREIWDQCnAVYNNTL NAVESKPLLFLHGILNECRNNFATKLRQDPSUVAKIDQIIKSQIYRFWVSEPYLKIGRSHT LYTHrrPDAVPQLPKECTlKHLSSYMEEKLKSMESKKNIESGKYEFDVDSSETDSTKDDG KPDDDDDDDDNFDDDDNFDDDTVEEEDASGDLFKNEKKDENKE _

19 SPELD (3D7) MCSTYSVEPLVYDSYEYVYLKPKKAGTSAVYYPLNISWKYVAKKRSVGCFGSRKKYTLIP EAYYYPYYYYYVYYYPSAARLVRTTKKEKVLKENNNKESEDENKQDNVGTEKKECDCSEK EKYIPTYVPLTESYYFPPSALYVPHYSVLVP _

20 PL (3D7) MSSILLFFWFQYLUSFSGT5YKRFWDGASIFLRNPYKITLGKSEKKGKVFSEFSEEEDSI VRRDTEKKKKWFGKGNDVNKKVKEKKNVILEEKQIGKEENNLVGQQNDSVGQQNDSVG QQNOSVGQQNDPVGQQNDSVGQQNDPVGQQNDPVGQQNDYVGQQNDSVGQQNDY VGQQNDYVGQQNDPVGQQNDSVGQQNDYVGQQNDPVGQQNDPVGQQNDPVGQQN DSVGQQNDYVGQQNDPVGQQNDYVGQQNDYVGQQNDPVGQQNDPVGQQNDPVEQ EDNVGIVKKEKKEKIIVEKENEIGVIERENISGVEV1NKDSEEDIYNSNVDKNFPADDNNR DBLVRRKKVSNKDSLFMNDIIRGENEDNDDDNEEEEEKVSKNLENVKLGELSKLQYIFG TNHDNDDDSEIRSESBDDIDELDEEEYDWTEHVYNYKPTTYLLPGLGGSTLIAEYKNAT IHSCSRYLLNSKPFRIWISLSRLLSlQSNIYCTFDTIRLKYDEKKNIYYNQPGVnDVEKFG NLKGIEYLDYFNNTGIGITKYFNWGQYFTSHGYVDGESIIGAPYDWRYPLSQQNYKILKE HIEYIYEKRNGTKVNUGHSLGGLYLNFFLSRWSKKWKQKHLSKIIFISTPFKGSVKnRA UQSRKDnSFRJTKLIKLSIPESMMKALGNSLGSLFDILPYREYYKRDQVVIUNMSNTPID EDHVQYLVRCGIYKPECYRNRADVNLKVYTLKNWHELLDDKLKAKYENYKLYRERYYNK DHGVPrYCLYSmNKKETEYLLYFETQNTREEPTIYYGTGDGnrVGTESLQACSNFYNTVLT HHFSDTSHVGILYTMETAKYIYDIVESPN _

21 E140 MVDFNDLSVELKKTEUKEDLRNLSHIINNEFSYFCQNENKNVSFNNNISSYYNDDIFSKS TLNNLYTSWKLEDFSHFDFSSILDILKRNQYVMCSIYFLLIFSCIYFLTLLLYTKCIKTTLKK WFCRYCSENINENNSNHNEQRTVLQNVINKSCYFITYSSIICLLLFLLLSGITYMHYFIKTK KGIHSNICNIYTRLDKFLLNKCLDPKKVDTSCYSAEHILNDLSSILEEYKKVKQQAKDDTLL DENTPFPLLERYnTFNKLNVLKNNINKNNTTLENEYFHTYPALKGISETLTTIISEGNKNF GNARNVIKEVKSTIKYSFHTVDETIRNVFKDSVPKlTGLrTQAGKSIKGINNKYKIKERlPK YTNIILLTNIILLLPPFLILLGIIIFMIFILMGYIQKNNNFFIKLFGHFSAYFGLLTIIILSFGILFL STSVIGGTSCILSERILKNELRFDILNNTLIDYCIKNESAPLIDDDITTSFVAKINSFDTGHI DHNINEYEKHFTILKESFFHKSLKFMDYIWIVIMKRENNTFLNRIRTEQVKKSLIJTGIINE NIKYENMEAIGIRSYLTTLNKIIFPENNGKICFNDIICEKENNTYNnENSiarTDQKYRNIR DGMDEHLRNDLDAIVQLFVYKARILKENIFDINDLDSNEKNKIGWSEYTPRNINGTQKKS IINTFLVNVIESINFSEIINFFDKMRDQFNVLKDULLKIDTLTENTKCNKLVKELINVRKDY CNNVVLNLSTLSVYUIFSrrSFLLWYLFLFLWFYYNIKPS _

22 AMA1 MRKLYCVLLLSAFEFTYMINFGRGQNYWEHPYQNSDVYRPINEHREHPKEYEYPLHQEH TYQQEDSGEDENTLQHAYPIDHEGAEPAPQEQNLFSSIEIVERSNYMGNPWTEYMAKYD IEEVHGSGIRVDLGEDAEVAGTQYRLPSGKCPVFGKGIIIENSNTTFLTPVATGNQYLKDG GFAFPPTEPLMSPMTLDEMRHFYKDNKYVKNLDELTLCSRHAGNMIPDNDKNSNYKYPA VYDDKDKKCHILYIAAQENNGPRYCNKDESKRNSMFCFRPAKDISFQNYTYLSKNWDN WEKVCPRKNLQNAKFGLWVDGNCEDIPHVNEFPAIDLFECNKLVFELSASDQPKQYEQH LTDYEKIKEGFKNKNASMIKSAFLPTGAFKADRYKSHGKGYNVVGNYNTETQKCEIFNVKP TCLINNSSYIATTALSHPIEVENNFPCSLYKDEIMKEIERESKRIKLNDNDDEGNKKIIAPRI FISDDKDSLKCPCDPEMVSNSTCRFFVCKCVERRAEVTSNNEVWKEEYKDEYADIPEHK PTYDKMKIIIASSAAVAVLATILMVYLYKRKGNAEKYDKMDEPQDYGKSNSRNDEMLDPE ASFWGEEKRASHTTPVLMEKPYY _

23 GARP MNVLFLSYNICILFFWCTLNFSTKCFSNGLLKNQNILNKSFDSITGRLLJNETELEKNKDDN SKSERLKEEKDEKDDVPTTSNDNLKNAHNNNEISSSTDPTNIINVNDKDNENSVDKKKD KKEKKHKKDKKEKKEKKDKKEKKDKKEKKHKKEKKHKKDKKKEENSEVMSLYKTGQHKP KNATEHGEENLYEEMVSEINNNAQGGLLLSSPYQYREQGGCGIISSVHETSNDTKDNDK ENISEDKKEDHQQEEMLKTLDKKERKQKEKEMKEQEKIEKKKKKQEEKEKKKQEKERKK QEKKERKQKEKEMKKQKKIEKERKKKEEKEKKKKKHDKENEETMQQPDQrSEETNNEIM VPLPSPLTDVTTPEEHKEGEHKEEEHKEGEHKEGEHKEEEHKEEEHKKEEHKSKEHKSKG KKDKGKKDKGKHKKAKKEKVKKHWKNVIEDEDKDGVEIINLEDKEACEEQHrTVESRPL SQPQCKLIDEPEQLTLMDKSKVEEKNLSIQEQLIGTIGRVNWPRRDNHKKKMAiaEEAEL QKQKHVDKEEDKKEESKEVEEESKEVQEDEEEVEEDEEEEEEEEEEEEEEEEEEEEEEDEV EEDEDDAEEDEDDAEEDEDDAEEDDDDAEEDDDDAEEDDDEDEDEDEEEEEDEEEEEE SEKKIKRNLRKNAKI _

24 PIESP2 MLLFFAKLWFTFFFWLLKYGKTRSYPKSGHKGHTKLNQPWRTLADFNDMFANQKNTF NFLKHINHYKNEQDTNNTHTPNHDEYSHNLPKNHEESNANMNNHNSFNDKSVNKKEAF DQFLQTLLNNYEIMHKEDESKESNQHNYKEGPSYEDKKNMYKEILKGYYNVFFENYAND TESNVHNKPEEVHKHEEIHKHRKLHKHEEVHKPEEFHKPEEFHKHEKVHKHEEVHKPEEV HKHEENHKHEENHKPQMVGQAPPEKEIRQESRTULGSFPQAGBLREDLWNKEDNKFS YALDPNDYASIEDKLLGSIFGYFKKNHDNLVKHLLQQIHTYKHKYMELKEQYINEVMKLK KIYNKSIMVIFIASCISILGPVMLHMHQNNPEEFFATILSFSISLGLHNLLLT _

25 SEA-l/SEP-1 MMENKYPNELFCYINRYNINEIIENGEEKYVNEYDEDKNMSINHMNENDGICEYEIPFLL DYVDDSNKEDSEKNSLKSYLDDGASULSKPDELENYNKQNENEFDENNNNKNNKIDQL KEKINIIIIPNKGVINNFEEILSMANRNDKNIEKKLNDRFYQICCKSIADINTHNLNKIKDLK KKKNNKGSLNIEHlDYGDlFLTIHDnKSNNKIKGNNKTNLLHDSSYEIKKKTRRGTNIYK NPFHHRGSYLTSYENQKDIIYLNNLNNIMMDKYSNCSDSRKKEYSHFNSQEFSYDKYSM KDRMFLKNLYMKQNRLRDKRGKYHKLGDYQNIENYRKTGEHSFDCMNMSDIMHSNKM SHVNIMDHM1YKDNNNMSKLVDTINSREKDVKNYDDNFESYNNFFKNNNDEQHICLEY DDTYNLKDTVKNIIVEEEQCGKGVAaCDKNEDVDDLFVSKKTNYSSNKKREDYEKVFLE DNLHLKQTPSKRTKINIIPDYYDNNRSNKSYKENEEDALFEVCGSLKNDDILYKDNKLNVI NEDNIKEEDDKESWHLDNDEDKKEEMYKDVYPNVLSCEKEnRRNEKYNKSLNSTSSFE KIDNPSEINVESKEDTEYFDLUKKYEDTKINVYDNESLLLDLSNELREEMAKGDSNKNVN KVEDNDNKKENICHDNIMEDICHNNNVEDMYRNNNVEDMYRNNNVEDMYRNNNVED MYRNNNVEDVCHNNNVEDVCHNNNVEDVCHNNNVEDVYHNNNVEDMYHDNNIEDVC n. Piasmodium polypeptide constructs

[0392] The present disclosure, among other things, utilizes RNA technologies as a modality to express one or more Plasmodium polypeptide construct that includes one or more malarial proteins, or one or more portions thereof, described herein. For example, In some embodiments, a Plasmodium polypeptide construct comprises one or more Plasmodium Rh5 invasion complex polypeptides or portions thereof (e.g., antigenic portions of one or more Plasmodium Rh5 Invasion complex polypeptides). In some embodiments, one or more polypeptides or portions thereof of a Plasmodium Rh5 invasion complex can Include one or more polypeptides or portions thereof of a Plasmodium reticulocyte-binding protein homolog 5 (Rh5), a Plasmodium Cysteine-Rjch Protective Antigen (CyRPA), a Plasmodium Rh5-lnteracting Protein (Rlpr), a Plasmodium P 113, a Plasmodium thrombospondin-related apical merozoite protein (TRAMP), and/or a Plasmodium cysteine-rich small secreted protein (CSS). In some embodiments, a portion of a Rh5 invasion complex polypeptide can be a characteristic portion of a Rh5 invasion complex polypeptide. In some embodiments, a Plasmodium polypeptide construct additionally Includes one or more additional amino acid sequences, such as a secretory signal (e.g., a heterologous secretory signal), a transmembrane region (e.g., a heterologous transmembrane region), a multimerization region, and/or a linker, as described herein.

A. Certain Plasmodium RhS Invasion Complex Polypeptides or Portions Thereof

1. RhS

[0393] In some embodiments, a Plasmodium polypeptide construct described herein includes one or more Plasmodium Rh5 invasion complex polypeptides or portions thereof (e.g., antigenic portions thereof) that comprise one or more Plasmodium reticulocyte-binding protein homolog 5 (Rh5) polypeptides or antigenic portions thereof. [0394] In some embodiments, a Plasmodium polypeptide construct described herein includes one or more polypeptides or antigenic portions of an Rh5 polypeptide, e.g., Plasmodium Rh5 polypeptide, e.g., P. fafciparum Rh5 (SEQ ID NO: 1), or a variant thereof (e.g., a glycosylation variant). In some embodiments, an Rh5 polypeptide may include a Plasmodium Rh5 N-terminal disordered domain, a first Plasmodium Rh5 ordered domain, a Plasmodium Rh5 linking disordered domain, a second Plasmodium Rh5 ordered domain (e.g., a C-terminal Plasmodium Rh5 ordered domain), or a combination thereof. A portion of Rh5 (or Rh5 polypeptide portion) (e.g., an antigenic portion of Rh5 or Rh5 antigenic portion) may refer to parts of an Rh5 polypeptide domain or parts spanning two or more Rh5 polypeptide domains. [0395] In some embodiments, a Plasmodium polypeptide construct comprises one or more antigenic portions of Rh5. In some embodiments, one or more antigenic portions of Rh5 comprise one or more Rh5 ordered domains. In some embodiments, a Plasmodium Rh5 polypeptide or antigenic portion thereof comprises 25, 30, 35, 40, or 45 contiguous amino adds of an Rh5 ordered domain, or a variant thereof (e.g., a glycosylation variant).

[0396] In some embodiments, a Plasmodium Rh5 antigenic portion comprises an Rh5 ordered domain that corresponds to amino adds 140-247 or amino adds 146-247 of a wild-type Rh5 (SEQ ID NO: 1) or variants thereof (e.g., glycosylation variants). In some embodiments, a Plasmodium Rh5 antigenic portion comprises an Rh5 ordered domain that corresponds to amino acids 297-526 of a wild-type Rh5 sequence (SEQ ID NO: 1), or a variant thereof (e.g., a glycosylation variant). In some embodiments, one or more antigenic portions of Rh5 comprise (i) an Rh5 ordered domain that corresponds to amino adds 140-247 or amino acids 146-247 of a wild-type Rh5 (SEQ ID NO: 1) or variants thereof and/or (ii) an Rh5 ordered domain that corresponds to amino adds 297-526 of a wild-type Rh5 sequence (SEQ ID NO: 1), or a variant thereof.

[0397] 140-247 Ordered Domain of RhS (SEQ ID NO. 118)

KNVNFLQYHFKELSNYNIANSIDILQEKEGHLDFVIIPHYTFLDYYKHLSYNSIYHKSSTYGKCIAVDAFIKKINETYDKVKSKCNDIKN DLIATIKKLEHPYDINNK

[0398] 146-247 Ordered Domain of RhS APMX (SEQ ID NO. 119)

QYHFKELSNYNIANSIDILQEKEGHLDFVIIPHYTFLDYYKHLSYNSIYHKSSTYGKCIAVDAFIKKINETYDKVKSKCNDIKNDLIATIK KLEHPYDINNK

[0399] 297-526 Ordered Domain of RhS (SEQ ID NO. 120)

NRTFKKMMDEYNTKKKKLIKCIKNHENDFNKICMDMKNYGTNLFEQLSCYNNNFCNTNGIRYHYDEYIHKLILSVKSKNLNKDLSDM TNILQQSELLLTNLNKKMGSYIYIDTIKFIHKEMKHIFNRIEYHTKIINDKTKIIQDKIKLNIWRTFQKDELLKRILDMSNEYSLFrTSDH LRQMLYNTFYSKEKHLNNIFHHL1YVLQMKFNDVPIKMEYFQTYKKNKPLTQ

[0400] In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise one or more unpaired cysteines. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise two unpaired cysteines, in some embodiments, one or more Plasmodium Rh5 antigenic portions comprise a cysteine at position 203, 329, or both, as numbered according to SEQ JD NO: 1. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise a tyrosine at position 203, 329, or both, as numbered according to SEQ ID NO: 1.

[0401] 140-247 Ordered Domain of RhS - C203Y (SEQ ID NO. 121)

KNVNFLQYHFKELSNYNIANSIDILQEKEGHLDFVIIPHYTFLDYYKHLSYNSIYHKSSTYGKYIAVDAFIKKINETYDKVKSKCNDIKN DLIATIKKLEHPYDINNK

[0402] 146-247 Ordered Domain of Rh5 APMX - C203Y (SEQ ID NO. 122)

QYHFKELSNYNIANSIDILQEKEGHLDFVIIPHYTFLDYYKHLSYNSIYHKSSTYGKYIAVDAFIKKINETYDKVKSKCNDIKNDLIATIK KLEHPYDINNK

[0403] 297-526 Ordered Domain of Rh5 - C329Y (SEQ ID NO. 123)

NRTFKKMMDEYNTKKKKLIKCIKNHENDFNKIYMDMKNYGTNLFEQLSCYNNNFCNTNGIRYHYDEYIHKULSVKSKNLNKDLSDM

TNILQQSELLLTNLNKKMGSYIYIDTIKFIHKEMKHIFNRIEYHTKIINDKTKIIQDKIKLNIWRTFQKDELLKRILDMSNEYSLFITSDH

LRQMLYNTFYSKEKHLNNIFHHUYVLQMKFNDVPIKMEYFQTYKKNKPLTQ

[0404] In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise one, two, three, or four N-linked glycosylation sites. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise two N-linked glycosylation sites. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise an amino acid substitution at one or more N-linked glycosylation sites, where the amino add substitution prevents glycosylation. In some embodiments, an amino acid substitution that prevents glycosylation comprises a NX[T/S] to QX[T/S] substitution. In some embodiments, amino acid substitution prevents glycosylation comprises a NX[T/S] to NXA substitution. In some embodiments, one or more Piasmocfium Rh5 antigenic portions comprise an amino add substitution at all of the N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation. [0405] In some embodiments, a Piasmodkim Rh5 antigenic portion comprises a Rh5 ordered domain that corresponds to amino adds 140-247 of a wild-type Rh5 (SEQ ID NO: 1) (or amino adds 140-247 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino add substitutions). In some embodiments, an Rh5 ordered domain corresponds to amino acids 140-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a cysteine at position 203. In some embodiments, an Rh5 ordered domain corresponds to amino adds 140-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a tyrosine at position 203. In some embodiments, an Rh5 ordered domain corresponds to amino adds 140-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises an N linked glycosylation site at N214. In some embodiments, an Rh5 ordered domain corresponds to amino adds 140-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises an amino add substitution that prevents glycosylation at position 214. In some embodiments, an amino add substitution that prevents glycosylation at position 214 comprises a NX[T/S] to QX[T/S] substitution. In some embodiments, amino add substitution prevents glycosylation at position 214 comprises a NX[T/S] to NXA substitution.

[0406] 140-247 Ordered Domain of RhS - N214Q (SEQ ID NO: 124)

KNVNFLQYHFKELSNYNIANSIDILQEKEGHLDFVIIPHYTFLDYYKHLSYNSIYHKSSTYGKCIAVDAFIKKIQETYDKVKSKCNDIKN

DUATIKKLEHPYDINNK

[0407] In some embodiments, a Rh5 ordered domain corresponds to amino adds 140-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a tyrosine at position 203 and glutamine at position 214.

[0408] 140-247 Ordered Domain of RhS - C203Y - N214Q (SEQ ID NO: 125)

KNVNFLQYHFKELSNYNIANSIDILQEKEGHLDFVIIPHYTFLDYYKHLSYNSIYHKSSTYGKYIAVDAFIKKIQETYDKVKSKCNDIKN

DUATIKKLEHPYDINNK

[0409] In some embodiments, an Rh5 antigenic portion (e.g., a Rh5 ordered domain) comprises a Plasmepsin X (PMX) deavage site. PMX is a conserved aspartic protease that is responsible for proteolytic processing and activation of proteins and other proteases involved in life cyde progression and/or development of Plasmodium.

[0410] In some embodiments, a PMX cleavage site of an Rh5 ordered domain comprises a sequence of FLQY (SEQ ID NO: 36), where cleavage occurs between L and Q. In some embodiments, a cleaved Rh5 ordered domain corresponds to amino adds 146-247 of a wild-type Rh5 (SEQ ID NO: 1) or a variant thereof.

In some embodiments, an Rh5 antigenic portion (e.g., an Rh5 ordered domain) does not comprise a Plasmepsin X (PMX) deavage site.

[0411] In some embodiments, a Plasmodium Rh5 antigenic portion comprises a Rh5 ordered domain that corresponds to amino adds 146-247 of a wild-type Rh5 (SEQ ID NO: 1) (or amino adds 146-247 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino add substitutions). In some embodiments, an Rh5 ordered domain corresponds to amino acids 146-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a cysteine at position 203. In some embodiments, an Rh5 ordered domain corresponds to amino adds 146-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a tyrosine at position 203. In some embodiments, an Rh5 ordered domain corresponds to amino adds 146-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises an N linked glycosylation site at N214. In some embodiments, an Rh5 ordered domain corresponds to amino adds 146-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises an amino add substitution that prevents glycosylation at position 214. In some embodiments, an amino add substitution that prevents glycosylation at position 214 comprises a NX[T/S] to QX[T/S] substitution. In some embodiments, amino add substitution prevents glycosylation at position 214 comprises a NX[T/S] to NXA substitution.

[0412] 146-247 Ordered Domain of RhS APMX - N214Q (SEQ ID NO: 126)

QYHFKELSNYNIANSIDILQEKEGHLDFVIIPHYTFLDYYKHLSYNSIYHKSSTYGKCIAVDAFIKKIQETYDKVKSKCNDIKNDLIATIK KLEHPYDINNK

[0413] In some embodiments, an Rh5 ordered domain corresponds to amino adds 146-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a tyrosine at position 203 and glutamine at position 214.

[0414] 146-247 Ordered Domain of RhS APMX - C203Y - N214Q (SEQ ID NO: 127)

QYHFKELSNYNIANSIDILQEKEGHLDFVIIPHYTFLDYYKHLSYNSIYHKSSTYGKYIAVDAFIKKIQETYDKVKSKCNDIKNDLIATIK KLEHPYDINNK

[0417] 297-526 Ordered Domain of RhS N297Q (SEQ ID NO. 128)

QRTFKKMMDEYNTKKKKLIKCIKNHENDFNKICMDMKNYGTNLFEQLSCYNNNFCNTNGIRYHYDEYIHKLILSVKSKNLNKDLSDM TNILQQSELLLTNLNKKMGSYIYIDTIKFIHKEMKHIFNRIEYHTKIINDKTKIIQDKIKLNIWRTFQKDELLKRILDMSNEYSLFITSDH LRQMLYNTFYSKEKHLNNIFHHUYVLQMKFNDVPIKMEYFQTYKKNKPLTQ

[0418] In some embodiments, an Rh5 ordered domain corresponds to amino adds 297-526 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a tyrosine at position 329 and/or glutamine at position 297.

[0419] In some embodiments, an Rh5 ordered domain corresponds to amino adds 297-526 of a wild-type Rh5 (SEQ ID NO: 1) and conyrises a cysteine at position 329 and asparagine at position 297.

[0420] 297-526 Ordered Domain of RhS - N297Q - C329Y (SEQ ID NO: 129)

QRTFKKMMDEYNTKKKKLIKQKNHENDFNKIYMDMKNYGTNLFEQLSCYNNNFCNTNGIRYHYDEYIHKLILSVKSKNLNKDLSDM TNILQQSELLLTNLNKKMGSYIYIDTIKFIHKEMKHIFNRIEYHTKHNDKTKIIQDKIKLNIWRTFQKDELLKRILDMSNEYSLFITSDH LRQMLYNTFYSKEKHLNNIFHHUYVLQMKFNDVPIKMEYFQTYKKNKPLTQ

[0421] In some embodiments, a Plasmodium Rh5 polypeptide or antigenic portion thereof comprises an Rh5 disordered domain or antigenic portion thereof. In some embodiments, an Rh5 disordered domain is an Rh5 N- terminal disordered domain that corresponds to amino adds 26-139 of a wild-type Rh5 (SEQ ID NO: 1). In some embodiments, an Rh5 disordered domain is an Rh5 linking disordered domain that corresponds to amino adds 248- 296 of a wild-type Rh5 (SEQ ID NO: 1).

[0422] 26-139 Disordered Domain of RhS (SEQ ID NO: 130)

ENAIKKTKNQENNLTLLPIKSrEEEKDDIKNGKDIKKBDNDKENIKTNNAKDHSTYIKSYLNTNVNDGI-KYLFIPSHNSFIKKYSVFN QINDGMLLNEKNDVKNNEDYKNVDY

[0423] 248-296 Disordered Domain of RhS (SEQ ID NO: 131)

NDDSYRYDISEHDDKSEETDDETEEVEDSIQDTDSNHTPSNKKKNDLM

[0424] In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise two Rh5 ordered domains and one Rh5 disordered domain. In some embodiments, one or more Plasmodium Rh5 antigenic portions correspond to amino adds 140-526 of a wild-type Rh5 (SEQ ID NO: 1). In some embodiments, one or more Piasmocfium Rh5 antigenic portions correspond to amino adds 26-247 and 297-526 of a wild-type Rh5 (SEQ ID NO: 1).

[0425] In some embodiments, a Plasmodium polypeptide construct does not include an Rh5 disordered domain. In some embodiments, a Plasmodium Rh5 polypeptide or antigenic portion thereof does not comprise an Rh5 disordered domain that corresponds to amino adds 26-139 or amino acids 248-296 of a wiki-type Rh5 (SEQ ID NO: 1).

In some embodiments, a Plasmodium polypeptide construct does not Indude an Rh5 secretory signal, e.g., does not indude a sequence corresponding to amino adds 1-25 of a wild-type Rh5 (SEQ ID NO: 1).

[0426] RhS secretory signal (SEQ ID NO: 132)

MIRIKKKULTIIYIHLFILNRLSF

[0427] In some embodiments, a Plasmodium polypeptide construct comprises one or more Plasmodium Rh5 polypeptides or antigenic portions thereof comprising two Rh5 ordered domains. In some embodiments, a Plasmodium polypeptide construct comprises two Plasmodium Rh5 antigenic portions, where each antigenic portion comprises an Rh5 ordered domain. In some embodiments, a Plasmodium polypeptide construct comprises two Rh5 ordered domains that correspond to (I) amino adds 140-247 or 146-247 of a wild-type Rh5 (SEQ ID NO: 1) or variants thereof and (II) amino adds 297-526 of a wild-type Rh5 (SEQ ID NO: 1) or variants thereof. In some embodiments, a Plasmodium polypeptide construct comprises two Rh5 ordered domains that are directly adjacent to one another.

[0428] In some embodiments, a Plasmodium polypeptide construct comprises an Rh5 polypeptide comprising exactly two antigenic portions of Rh5, where the two antigenic portions comprise or consist of two Rh5 ordered domains. In some embodiments, two Rh5 ordered domains correspond to (i) amino acids 140-247 or amino adds 146-247 of a wild-type Rh5 (SEQ ID NO: 1) or variants thereof and (II) amino acids 297-526 of a wild-type Rh5 (SEQ ID NO: 1) or variants thereof.

[0429] In some embodiments, a Plasmodium polypeptide construct comprising two Rh5 ordered domains, where a first Rh5 ordered domain corresponds to amino adds 140-247 or amino acids 146-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a tyrosine at position 203 and glutamine at position 214, and where a second Rh5 ordered domain corresponds to amino adds 297-526 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a tyrosine at position 329 and/or glutamine at position 297.

[0430] In some embodiments, a Plasmodium polypeptide construct comprising two Rh5 ordered domains, where a first Rh5 ordered domain corresponds to amino acids 140-247 or amino acids 146-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a tyrosine at position 203 and glutamine at position 214, and where a second Rh5 ordered domain corresponds to amino adds 297-526 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a cysteine at position 329 and asparagine at position 297.

[0431] In some embodiments, a Plasmodium polypeptide construct does not include a Rh5 disordered domain (e.g., does not indude amino adds 26-139 or amino adds 248-296 of a wild-type Rh5 (SEQ ID NO: 1). In some embodiments, a Plasmodium polypeptide construct does not include a Rh5 secretory signal, e.g., does not indude a sequence corresponding to amino adds 1-25 of a wild-type Rh5 (SEQ ID NO: 1).

[0432] In some embodiments, a Plasmodium polypeptide construct comprises one or more antigenic portions of Plasmodium Rh5 comprising a PMX deavage site. In some embodiments, a Plasmodium polypeptide construct comprises one or more antigenic portions of Plasmodium Rh5 comprising a PMX cleavage site that comprises or consists of an amino add sequence of NFLQ (SEQ ID NO: 189).

2. CyRPA [0433] In some embodiments, a Plasmodium polypeptide construct described herein Includes one or more Plasmodium Rh5 invasion complex polypeptides or portions thereof (e.g., antigenic portions thereof) that comprise one or more Plasmodium Cysteine-Rich Protective Antigen (CyRPA) polypeptides or antigenic portions thereof. [0434] In some embodiments, a Plasmodium polypeptide construct described herein includes one or more regions or portions (e.g., antigenic portions) of a CyRPA, e.g., Plasmodium CyRPA, e.g., P. falciparum CyRPA (SEQ ID NO: 3), or a variant thereof (e.g., one or more antigenic portions of a CyRPA, e.g., Plasmodium CyRPA, e.g., P. falciparum CyRPA).

[0435] In some embodiments, a Plasmodium polypeptide construct comprises an antigenic portion of CyRPA. In some embodiments, a Plasmodium CyRPA polypeptide or antigenic portion thereof comprises 25, 30, 35, 40, or 45 contiguous amino adds of CyRPA, or a variant thereof (e.g., a glycosylation variant).

[0436] In some embodiments, a Plasmodium CyRPA antigenic portion does not comprise an endogenous signal sequence. In some embodiments, a Plasmodium CyRPA antigenic portion is operably linked to a heterologous signal sequence, as further described herein. In some embodiments, a Plasmodium CyRPA antigenic portion comprises or consists of amino adds 30-362 of a wild-type CyRPA (SEQ ID NO: 3) or a variant thereof (e.g., a glycosylation variant). In some embedments, a Plasmodium CyRPA antigenic portion comprises or consists of a sequence according to SEQ ID NO: 133 or a variant thereof (e.g., a glycosylation variant). In some embodiments, a Plasmodium CyRPA antigenic portion comprises or consists of a sequence with at least 85% sequence identity to SEQ ID NO: 133 or a variant thereof (e.g., a glycosylation variant).

[0437] 30-362 portion of CyRPA (SEQ ID NO. 133)

SRHVFIRTELSFIKNNVPCIRDMFFIYKRELYNICLDDLKGEEDETHIYVQKKVKDSWmLNDLFKETDLTGRPHIFAYVDVEEIIILLCE DEEFSNRKKDMTCHRFYSNDGKEYNNSEmSDYILKDKLLSSYVSLPLXIENREYFLICGVSPYKFKDDNKKDDILCMASHDKGETW

GTKP/IKYDNYKLGVQYFFLRPYISKNDLSFHFYVGDNINNVKNVNFIECTHEKDLEFVCSNRDFLKDNKVLQDVSRNDEYIVSYGN DNNFAECYIFFNNENSIUKPEKYGNTTAGCYGGTFVKIDENRTLFIYSSSQGIYNIHnYYANYE

[0438] In some embodiments, a Plasmodium CyRPA antigenic portion comprises or consists of amino acids 29- 362 of a wild-type CyRPA (SEQ ID NO: 3) or a variant thereof (e.g., a glycosylation variant). In some embodiments, a Plasmodium CyRPA antigenic portion comprises or consists of a sequence according to SEQ ID NO: 169 or a variant thereof (e.g., a glycosylation variant). In some embodiments, a Plasmodium CyRPA antigenic portion comprises or consists of a sequence with at least 85% sequence identity to SEQ ID NO: 169 or a variant thereof (e.g., a glycosylation variant).

[0439] 29-362 portion of CyRPA (SEQ ID NO. 169)

DSRHVFIRTELSFIKNNVPCIRDMFFIYKRELYNICLDDLKGEEDETHIYVQKKVKDSWrTLNDLFKETDLTGRPHIFAYVDVEEIIILLC EDEEFSNRKKDMTCHRFYSNDGKEYNNSEItISDYILKDKLLSSYVSLPLKIENREYFUCGVSPYKFKDDNKKDDILCMASHDKGET

WGTKIVIKYDNYKLGVQYFFLRPYISKNDLSFHFYVGDNINNVKNVNFIECTHEKDLEFVCSNRDFLKDNKVLQDVSTLNDEYIVSYG NDNNFAECYIFFNNENSILIKPEKYGNTTAGCYGGTFVKIDENRTLFIYSSSQGIYNIHTIYYANYE

[0440] In some embodiments, one or more Plasmodium CyRPA antigenic portions comprise one or more N- llnked glycosylation sites. In some embodiments, a Plasmodium CyRPA antigenic portion comprises one, two, or three N-linked glycosylation sites. In some embodiments, a Plasmodium CyRPA antigenic portion comprises an amino acid substitution at one or more N-linked glycosylation sites, where the amino acid substitution prevents glycosylation. In some embodiments, an amino acid substitution that prevents glycosylation comprises a NX[T/S] to QX[T/S] substitution. In some embodiments, amino acid substitution prevents glycosylation comprises a NX[T/S) to NXA substitution. In some embodiments, a Plasmodium CyRPA antigenic portion comprises two amino acid substitutions at N-linked glycosylation sites that prevent glycosylation. In some embodiments, a Plasmodium CyRPA antigenic portion comprises an amino acid substitution at all of the N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation.

[0441] In some embodiments, a Plasmodium CyRPA antigenic portion comprises an asparagine at position 145, position 322, position 338, or a combination thereof, as numbered according to SEQ ID NO: 133. In some embodiments, a Plasmodium CyRPA antigenic portion comprises a glutamine at position 145, position 322, position

338, or a combination thereof, as numbered according to SEQ ID NO: 3. In some embodiments, a Plasmodium CyRPA antigenic portion comprises a glutamine at position 145, position 322, and position 338, as numbered according to SEQ ID NO: 3. In some embodiments, a Plasmodium CyRPA antigenic portion comprises or consists of an amino add sequence of SEQ ID NO: 134.

[0442] In some embodiments, a Plasmodium CyRPA antigenic portion comprises an asparagine at position 146, position 323, position 339, or a combination thereof, as numbered according to SEQ ID NO: 3. In some embodiments, a Plasmodium CyRPA antigenic portion comprises a glutamine at position 146, position 323, position

339, or a combination thereof, as numbered according to SEQ ID NO: 3. In some embodiments, a Plasmodium CyRPA antigenic portion comprises a glutamine at position 146, position 323, and position 339, as numbered according to SEQ ID NO: 3. In some embodiments, a Plasmodium CyRPA antigenic portion comprises or consists of an amino add sequence of SEQ ID NO: 170.

[0443] 30-362 portion of CyRPA Aolvcan (SEQ ID NO. 134)

SRHVFIRTELSFIKNNVPCIRDMFFIYKRELYNICLDDLKGEEDETHIYVQKKVKDSWITLNDLFKETDLTGRPHIFAYVDVEEIIILLCE DEEFSNRKKDMTCHRFYSNDGKEYONSEITISDYILKDKLLSSYVSLPLKIENREYFLICGVSPYKFKDDNKKDDILCMASHDKGETW GTKMKYDNYKLGVQYFFLRPYISKNDLSFHFYVGDNINNVKNVNFIECTHEKDLEFVCSNRDFLKDNKVLQDVSTLNDEYIVSYGN DNNFAECYIFFNNENSILIKPEKYGOTTAGCYGGTFVKIDEORTLFIYSSSQGIYNIHTIYYANYE

[0444] 29-362 portion of CvRPA Aolvcan f SEO ID NO. 1701

DSRHVFIRTELSFIKNNVPCIRDMFFIYKRELYNICLDDLKGEEDETHIYVQKKVKDSWnLNDLFKETDLTGRPHIFAYVDVEEIIILLC EDEEFSNRKKDMTCHRFYSNDGKEYONSEmSDYILKDKLLSSYVSLPLKIENREYFUCGVSPYKFKDDNKKDDILCMASHDKGET WGTKIVIKYDNYKLGVQYFFLRPYISKNDLSFHFYVGDNINNVKNVNFIECrHEKDLEFVCSNRDFLKDNKVLQDVSTLNDEYIVSYG

NDNNFAECYIFFNNENSILIKPEKYGQTIAGCYGGTFVKIDEQBILFIYSSSQGIYNIHnYYANYE

3. P113

[0445] In some embodiments, a Plasmodium polypeptide construct described herein Includes one or more Plasmodium Rh5 invasion complex polypeptides or portions thereof (e.g., antigenic portions thereof) that comprise one or more Plasmodium P113 polypeptides or antigenic portions thereof.

[0446] In some embodiments, a Plasmodium polypeptide construct described herein includes one or more regions or portions (e.g., antigenic portions) of a P113, e.g., Plasmodium P113, e.g., P. falciparum P113 (SEQ ID NO: 6), or a variant thereof (e.g., one or more antigenic portions of a P113, e.g., Plasmodium P113, e.g., P. falciparum P113).

[0447] In some embodiments, a Plasmodium polypeptide construct comprises one or more antigenic portion of P113. In some embodiments, a Hasmodum P113 polypeptide or antigenic portion thereof comprises 25, 30, 35, 40, or 45 contiguous amino acids of a Pl 13 polypeptide, or a variant thereof (e.g., a glycosylation variant). In some embodiments, a Plasmodium polypeptide construct comprises an antigenic portion of P113.

[0448] In some embodiments, a Plasmodium P113 antigenic portion does not comprise an endogenous signal sequence. In some embodiments, a Plasmodium P113 antigenic portion is operably linked to a heterologous signal sequence, as described further herein. In some embodiments, a Plasmodium Pl 13 antigenic portion does not comprise a GPI anchor site. In some embodiments, a Plasmodium P113 antigenic portion comprises or consists of amino acids 23-939 of a wild-type P113 (SEQ ID NO: 6) or a variant thereof (e.g., a glycosylation variant). In some embodiments, a Plasmodium P113 antigenic portion comprises or consists of a sequence according to SEQ ID NO: 135. In some embodiments, a Plasmodium P113 antigenic portion comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 135.

[0449] 23-939 portion of P113 (SEQ ID NO: 135)

YVHNDVIKFGEENSLKCSQGNLYVLHCEVQCLNGNNEIIHKRCNDDIEKKCNGNNKCIYFFEYELRKKTQSFRNKNSIEISECVESEQ NEVKTSTTCLLSNSFILDEAFIQYFFFIKNKNEEPVICKDGNINIKSALLHSPFCEIKLKDISEYIRKKCDNNKECLIDPLDVQKNLLNEE DPCYINNSYVSVNWCNKEEEIGDESTDSSSMEIQDSTSNEQDENVKGMSSSQEMNSNNDENKNQDNESDDDVNNNNNNNNDD QDEQGNDGDVTSSMNKNEDNKDLEHGSSNDVNNNTDTLVNNKENKEFVLKEKSSLTSKINKELAHRTALFNKLADNISLLLNKKYD SFEIKDVLEDRYNEMKRDANPDVYYIYLMDTLDIEKIEDINLEEVKMSLLASLKETMNKIDnEKKIEEFKNKYISLYNKVKTTMPELFD

LNEDLVLLYNDFPFDNGMISSDIFFKYNPSENIMDHQEMVKKGSITEDELRIVNDLEPLDNYRRRKRrTELRKILVEKLRILYLEKNNLF NTQASCIKSYCYKNPLNLKTLEVLLKKNYYRLKENKDYDWSSIIQHLDNVDANKKKKWLTHERILKKLQVLIAEGYKRINEKEKDIDR RMAVYNALYEKAQSYNLQKLFNDSNDFLKKYAIMGNSFDDGDEVFGSQSSNFNIFDSNNTDQNNEQEQPKQDDQLLNNNNDDVLS ESNNENKEKTSDDATHKETQEKSDQEPSQN1QEDNSDEKHAENEENVEQIETDSNVSEEANDENKDNMQTTTDEGTEELQQNDE DAESLTKENSKSEEQENEDSTDAEAIDKEEVETEEKGKDEQKKDEQKEQDEEEDGEKENKHKSSETTNETVTDIEENKNEVKGEEHL QGSEQSIEASESSQKDETKETEDKEEYVNANDDESSEEDTTPNE

[0450] In some embodiments, a Plasmodium P113 antigenic portion comprises one or more N-linked glycosylation sites. In some embodiments, a Plasmodium P113 antigenic portion comprises one, two, three, four, five, six, seven, or eight N-linked glycosylation sites. In some embodiments, a PlasnxxSum P113 antigenic portion comprises an amino acid substitution at one or more N-llnked glycosylation sites, where the amino add substitution prevents glycosylation. In some embodiments, an amino add substitution that prevents glycosylation comprises a NX[T/S] to QX[T/S] substitution. In some embodiments, amino add substitution prevents glycosylation comprises a NX(T/S] to NXA substitution. In some embodiments, a Plasmodium P113 antigenic portion comprises two, three, four, five, six, or seven amino acid substitutions at N-linked glycosylation sites that prevent glycosylation. In some embodiments, a Plasmodium P113 antigenic portion comprises an amino add substitution at all of the N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation.

[0451] In some embodiments, a Plasmodium P113 antigenic portion comprises an asparagine at position 207, position 268, position 317, position 360, position 661, position 697, position 779, position 876, or a combination thereof, as numbered according to SEQ ID NO: 6. In some embodiments, a Plasmodium P113 antigenic portion comprises a glutamine at position 207, position 268, position 317, position 360, position 661, position 697, position 779, position 876, or a combination thereof, as numbered according to SEQ ID NO: 6. In some embodiments, a Plasmodium P113 antigenic portion comprises a glutamine at position 207, position 268, position 317, position 360, position 661, position 697, position 779, and position 854, as numbered according to SEQ ID NO: 6. In some embodiments, a Plasmodium P113 antigenic portion comprises or consists of an amino acid sequence of SEQ ID NO: 136.

[0452] 23-939 portion of P113 Aolvcan (SEQ ID NO. 136)

YVHNDVIKFGEENSLKCSQGNLYVLHCEVQCLNGNNEIIHKRCNDDIEKKCNGNNKCIYFFEYELRKKTQSFRNKNSIEISECVESEQ NEVKTSTTCLLSNSFILDEAFIQYFFFIKNKNEEPVICKDGNINIKSALLHSPFCEIKLKDISEYIRKKCDNNKECLIDPLDVQKNLLNEE DPCYIQHSYVSVNWCNKEEEIGDESTDSSSMEIQDSTSNEQDENVKGMSSSQEMNSNNDENKNQDQESDDDVNNNNNNNNDD QDEQGNDGDVTSSMNKNEDNKDLEHGSSNDVNQtnPTLVNNKENKEFVLKEKSSLTSKINKELAHRTALFNKLADQI§LLLNKKYD SFEIKDVLEDRYNEMKRDANPDVYYIYLMDRDIEKIEDINLEEVKMSLLASLKETMNKIDTIEKKIEEFKNKYISLYNKVKTTMPELFD

LNEDLVLLYNDFPFDNGMISSDIFFKYNPSENIMDHQEMVKKGSnEDELRIVNDLEPLDNYRRRKRITHRKILVEKLRILYLEKNNLF NTQASCIKSYCYKNPLNLICrLEVLIKKNYYRLKENKDYDWSSIIQHLDNVDANKKKKWLTHERILKKLQVLIAEGYKRINEKEKDIDR RMAVYNALYEKAQSYNLOKLFODSNDFLKKYAIMGNSFDDGDEVFGSOSSNFNIFDSONTDONNEOEOPKODDOLLNNNNDDVL SESNNENKEKTSDDATHKETQEKSDQEPSQNIQEDNSDEKHAENEENVEQIETDSQYSEEANDENKDNMQTTTDEGTEELQQNDE DAESLTKENSKSEEQENEDSTDAEAIDKEEVETEEKGKDEQKKDEQKEQDEEEDGEKENKHKSSETTQByTDIEENKNEVKGEEHL QGSEQSIEASESSQKDETKETEDKEEYVNANDDESSEEDTTPNE 4. Ripr [0453] In some embodiments, a Plasmodium polypeptide construct described herein includes one or more Plasmodium Rh5 invasion complex polypeptides or portions thereof (e.g., antigenic portions thereof) that comprises one or more Plasmodium Rh5-interacting Protein (Ripr) polypeptides or antigenic portions thereof.

[0454] In some embodiments, a Plasmodium polypeptide construct described herein includes one or more regions or portions (e.g., antigenic portions) of a Ripr, e.g., Plasmodium Ripr, e.g., P. falciparum Ripr (SEQ ID NO: 2), or a variant thereof (e.g., one or more antigenic portions of a Ripr, e.g., Plasmodium Ripr, e.g., P. falciparum Ripr). In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 178. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 178. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 177. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 177. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 176. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 176. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide comprises or consists of an amino acid sequence according to SEQ ID NO: 175. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 175. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 174. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide comprises or consists of an amino add sequence with at least 85% sequence Identity to an amino acid sequence according to SEQ ID NO: 174. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 173. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 173. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 172. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 172. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 171. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 171.

[0455] 20-1086 portion of Ripr (SEQ ID NO: 171)

IDUEGIFYEKNBDKLTFSLDHRVRDNlKTDLILNNNGENDYAYLNKYVYnLNRDSTEKIKTFFSHNKDMKSCDYFISKEYNSSDKT

NQICYKKTECGWlPNSEBKTNKrTNDKLYCAHFNSTBlIIYYISQPLLLEPHVVYEETFFEKGKNDQINCQGMYISLRSVHVHTHNAI LQQETL1YIKNLCDGKNNCKFDFDSIKYENKSLTHYLFFINIQYQCISPLNLQENEMCDVYNDDTHKATCKYGFNKIELLKNVCEENY RCTQDICSVNQFCDGENETUTCKTSLLPSAKNNCEYNDLCTVLNCPENSTCEQIGNGKKAECKCENGKYYHNNKCYTKNDLELAIKI EPHKKEKFYKNNLYQGKALKPEYJFMQCENGFSIEVINAYVSCYRVSFNLNKLKYVTESLKKMCDGKTKCAYGNTIDPIDDLNHHNIC NNFN7IFKYDYLCVFNNQNTTSDKNSHLHSNIPSLYNSSILPDINKSKFHLISRNSRTNQYPHNNISMLBQNBSSHNSNQFSTDPHT NSNNINNMNIKKVBFRSRFSSKLQCQGGKINIDKAILKGGEGCNDLLLTNSLKSYCNDLSECDIGLIYHFDTYCINDQYLFVSYSCSN LCNKCHNNSTCYGNRFNYDCFCDNPYISKYGNKLCERPNDCESVLCSQNQVCQILPNDKLICQCEEGYKNVKGKCVPDNKCDLSCPS NKVCVIENGKQTCKCSERFVLENGVCICANDYKMEDGINCIAKNKCKRKEYENICTNPNEMCAYNEETDIVKCECKEHYYRSSRGECI LNDYCKDINCKENEECSIVNFKPECVCKENLKKNNKGECIYENSCLINEGNCPKDSKCIYREYKPHECVCNKQGHVAVNGKCVLEDKC VHNKKCSENSICVNVMNKEPICVCTYNYYKKDGVCLIQNPCLXDNGGCSRNSECTFKYSKINCTCKENYKNKDDSCVPNTNEYDESF TFQYNDDASIILGACGMIEFSYTYNQIIWKINNSKESYVFYYDYPTAGNIEVQIKNEIFHTIIYLKKKIGNSVIYDDFQVDHQTCIYENV FYYSNQN

[0456] 20-1086 portion of Ripr Aolvcan (SEQ ID NO: 172)

IDLIEGIFYEKNBDKLTFSLDHRVRDNLKTDLILNNNGENDYAYLNKYVYnLNRDSTEKIKTFFSHNKDMKSCDYFISKEYQSSDICr NQICYKKTFCGVVIPNSEBKrNKrTNDKLYCAHFQSTHIIIYYISQPLLLEPHVVYEETFFEKGKNDQINCQGMYISLRSVHVHTHNAI LQQETLTYIKNLCDGKNNCKFDFDSIKYEQKSLTHYLFFINIQYQCISPLNLQENEMCDVYNDDTHKATCKYGFNKIELLKNVCEENY RCTQDICSVNQFCDGEQEnnaCTSLLPSAKNNCEYNDLCTVLNCPEQSTCEQIGNGKKAECKCENGKYYHNNKCYTKNDLELAIKI EPHKKEKFYKNNLYQGKALKPEYIFMQCENGFSIEVINAYVSCYRVSFNLNKLKYVTESLKKMCDGKTKCAYGNTIDPIDDLNHHNIC NNFNTIFKYDYLCVFNNQQrTSDKNSHLHSNIPSLYQSSILPDIQKSKFHLISRNSRTNQYPHNQISMLEIQNHSSHNSNQFSTDPHT NSNNINNMNIKKVBFRSRFSSKLQCQGGKINIDKAILKGGEGCNDLLLTNSLKSYCNDLSECDIGLIYHFDTYCINDQYLFVSYSCSN LCNKCHQNSTCYGNRFNYDCFCDNPYISKYGNKLCERPNDCESVLCSQNQVCQILPNDKUCQCEEGYKNVKGKCVPDNKCDLSCPS NKVCVIENGKQTCKCSERFVLENGVCICANDYKMEDGINCIAKNKCKRKEYENICTNPNEMCAYNEETDIVKCECKEHYYRSSRGECI LNDYCKDINCKENEECSIVNFKPECVCKENLKKNNKGECIYENSCLINEGNCPKDSKCIYREYKPHECVCNKQGHVAVNGKCVLEDKC VHNKKCSENSICVNVMNKEPICVCTYNYYKKDGVCLIQNPCLKDNGGCSRNSECTFKYSKIQCTCKENYKNKDDSCVPNTNEYDESF TFQYNDDASIILGACGMIEFSYIYNQIIWKIQNSKESYVFYYDYPTAGNIEVQIKNEIFFrniYLKKKIGNSVIYDDFQVDHQTGIYENV FYYSNQN

[0457] 530-1086 portion of Ripr Aolvcan (SEQ ID NO: 173)

LEIQNEISSHNSNQFSTDPHTNSNNINNMNIKKVEIFRSRFSSKLQCQGGKINIDKAILKGGEGCNDLLLTNSLKSYCNDLSECDIGLI YHFDTYCINDQYLFVSYSCSNLCNKCHQNSTCYGNRFNYDCFCDNPYISKYGNKLCERPNDCESVLCSQNQVCQILPNDKUCQCEE GYKNVKGKCVPDNKCDLSCPSNKVCVIENGKQTCKCSERFVLENGVCICANDYKMEDGINCIAKNKCKRKEYENICTNPNEMCAYNE ETDIVKCECKEHYYRSSRGECILNDYCKDINCKENEECSIVNFKPECVCKENLKKNNKGECIYENSCUNEGNCPKDSKCIYREYKPHE

CVCNKQGHVAVNGKCVLEDKCVHNKKCSENSICVNVMNKEPICVCTYNYYKKDGVCLIQNPCLKDNGGCSRNSECTFKYSKIQCTC KENYKNKDDSCVPNTNEYDESFTFQYNDDASIILGACGMIEFSYIYNQIIWKIQNSKESYVFYYDYPTAGNIEVQIKNEIFHTIIYLKKK IGNSVIYDDFQVDHQTCIYENVFYYSNQN

[0458] 720-934 portion of Ripr (SEQ ID NO: 174)

DLSCPSNKVCVIENGKQTCKCSERFVLENGVCICANDYKMEDGINCIAKNKCKRKEYENICTNPNEMCAYNEErDIVKCECKEHYYRS SRGECILNDYCKDINCKENEECSIVNFKPECVCKENLKKNNKGECIYENSCLINEGNCPKDSKCIYREYKPHECVCNKQGHVAVNGKC VLEDKCVHNKKCSENSICVNVMNKEPICVCTYNYYKKDGVCUQ [0459] 769-900 portion of Ripr (SEQ ID NO: 175)

KNKCKRKEYENICTNPNEMCAYNEETDIVKCECKEHYYRSSRGECILNDYCKDINCKENEECSIVNFKPECVCKENLKKNNKGECrYE

NSCUNEGNCPKDSKCIYREYKPHECVCNKQGHVAVNGKCVLED

[0460] 769-856 portion of Ripr (SEQ ID NO: 176)

KNKCKRKEYEN1CTNPNEMCAYNEETDIVKCECKEHYYRSSRGECILNDYCKDINCKENEECSIVNFKPECVCKENLKKNNKGECIYE

[0461] 817-900 portion of Ripr (SEQ ID NO: 177)

DYCKDINCKENEECSIVNFKPECVCKENLKKNNKGECIYENSCLINEGNCPKDSKCIYREYKPHECVCNKQGHVAVNGKCVLED

[0462] 817-856 portion of Ripr (SEQ ID NO: 178) DYCKDINCKENEECSIVNFKPECVCKENLKKNNKGECIYE

[0463] In some embodiments, a Plasmodium polypeptide construct comprises one or more antigenic portions of Ripr. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide comprises 25, 30, 35, 40, or 45 contiguous amino acids of a Ripr polypeptide, or a variant thereof (e.g., a glycosylation variant). In some embodiments, a Plasmodium polypeptide construct comprises one or more antigenic portions of Ripr.

[0464] In some embodiments, an Plasmodium Ripr antigenic portion comprises one or more N-linked glycosylation sites. In some embodiments, a Plasmodium Ripr antigenic portion comprises one, two, three, four, five, six, seven, eight, nine, ten, eleven, or twelve N-linked glycosylation sites. In some embodiments, a Plasmodium Ripr antigenic portion comprises an amino acid substitution at one or more N-linked glycosylation sites, wherein the amino acid substitution prevents glycosylation. In some embodiments, an amino acid substitution that prevents glycosylation comprises a NX[T/S] to QX[T/S] substitution. In some embodiments, amino acid substitution prevents glycosylation comprises a NX[T/S] to NXA substitution. In some embodiments, a Plasmodium Ripr antigenic portion comprises an amino acid substitution at all of the N-linked glycosylation sites, wherein the amino acid substitution prevents glycosylation.

[0465] In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide comprises an asparagine at position 646, position 964, position 1021, or a combination thereof, as numbered according to SEQ ID NO: 2. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide comprises a glutamine at position 646, position 964, position 1021, or a combination thereof, as numbered according to SEQ ID NO: 2. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide comprises a glutamine at position 646, position 964, and position 1021, as numbered according to SEQ ID NO: 2. In some embodiments, the antigenic portion of a Plasmodium Ripr comprises or consists of an amino add sequence according to SEQ ID NO: 173.

[0466] 530-1086 portion of Ripr (SEQ ID NO: 190)

LEIQNEISSHNSNQFSTDPHTNSNNINNMNIKKVEIFRSRFSSKLQCQGGKINIDKAILKGGEGCNDLLLTNSLKSYCNDLSECDIGLI YHFDTYCINDQYLFVSYSCSNLCNKCHNNSTCYGNRFNYDCFCDNPYISKYGNKLCERPNDCESVLCSQNQVCQILPNDKLICQCEE GYKNVKGKCVPDNKCDLSCPSNKVCVIENGKQTCKCSERFVLENGVCICANDYKMEDGINCIAKNKCKRKEYENICTNPNEMCAYNE ETDIVKCECKEHYYRSSRGECILNDYCKDINCKENEECSIVNFKPECVCKENLKKNNKGECIYENSCLINEGNCPKDSKCIYREYKPHE

CVCNKQGHVAVNGKCVLEDKCVHNKKCSENSICVNVMNKEPICVCnrNYYKKDGVCLIQNPCLKDNGGCSRNSECTFKYSKINCTCK ENYKNKDDSCVPNTNEYDESFTFQYNDDASIILGACGMIEFSYIYNQIIWKINNSKESYVFYYDYPTAGNIEVQIKNEIFHTIIYLKKKI GNSVIYDDFQVDHQTCIYENVFYYSNQN

[0467] In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide comprises an asparagine at position 103, position 144, position 228, position 303, position 334, position 480, position 498, position 506, position 526, position 646, position 964, position 1021, or a combination thereof, as numbered accordng to SEQ ID NO: 2. In some embodiments, an antigenic portion of a Pfasmodium Ripr polypeptide comprises a ^utamine at position 103, position 144, position 228, position 303, position 334, position 480, position 498, position 506, position 526, position 646, position 964, position 1021, or a combination thereof, as numbered according to SEQ ID NO: 2. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide comprises a glutamine at position 103, position 144, position 228, position 303, position 334, position 480, position 498, position 506, position 526, position 646, position 964, and position 1021, as numbered according to SEQ ID NO: 2. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide comprises or consists of SEQ ID NO: 172.

[0468] In some embodiments, a Plasmodium Ripr antigenic portion comprises a PMX cleavage site. In some embodiments, a PMX cleavage site of a Ripr antigenic portion comprises a sequence of GNISMLEIQNEE (SEQ ID NO: 37). In some embodiments, a Plasmodium Ripr antigenic portion does not comprise a PMX cleavage site. In some embodiments, a PMX cleavage site of a Ripr antigenic portion comprises or consists of a sequence of SMLE (SEQ ID NO: 191).

5. TRAMP some embodiments, an antigenic portion of a Plasmodium TRAMP polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 179. In some embodiments, an antigenic portion of a Plasmodium TRAMP polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 179.

[0471] 42-352 portion of TRAMP (SEQ ID NO: 179)

LQVYQNNGNFKMEILEPECT1QKNLPSEEEKEEFIDFDRKELLLHDFWIKNSQFTDEKTLELYLSKEENNKSYLMLTFYLGDLKLMIG HNSPYEISLILNVATVNKNMSNCKNNSYNIVLLIGTDVFNTSDLEILEGPIQFSLGKSSGAFRINVTNFFLNNTWKAFTKDKISFLIKPE ENCYTILENKINKPQLLIEKICrTFYSEWGEWSNCSMDCDHPDNVQIRERECIHPSGDCFKGDLKESRPCIPLPPCNELFSHKDNSTF KILMIILPIVLVISimLYHIFYKRKGAEKELYENVAGRYMYD

[0472] In some embodiments, an antigenic portion of a Plasmodium TRAMP polypeptide comprises 25, 30, 35, 40, or 45 contiguous amino acids of a TRAMP polypeptide, or a variant thereof (e.g., a glycosylation variant). In some embodiments, a Plasmodium polypeptide construct comprises an antigenic portion of TRAMP.

[0473] In some embodiments, a Plasmodium TRAMP antigenic portion comprises one or more N-linked glycosylation sites. In some embodiments, a Plasmodium TRAMP antigenic portion comprises one, two, three, four, five, six, seven, or eight N-linked glycosylation sites. In some embodiments, a PlasmodiumTRMAP antigenic portion comprises an amino acid substitution at one or more N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation. In some embodiments, an amino add substitution that prevents glycosylation comprises a NX[T/S] to QX[T/S] substitutlon. In some embodiments, amino add substitution prevents glycosylation comprises a NX[T/S] to NXA substitution. In some embodiments, a Plasmodium TRAMP antigenic portion comprises an amino add substitution at all of the N-linked glycosylation sites, wherein the amino acid substitution prevents glycosylation. [0474] In some embodiments, an antigenic portion of a Plasmodium TRAMP polypeptidecomprises an asparagine at position 149, position 195, position 202, or a combination thereof, as numbered according to SEQ ID NO: 4. In some embodiments, an antigenic portion of a Plasmodium TRAMP polypeptide comprises a glutamine at position 149, position 195, position 202, or a combination thereof, as numbered according to SEQ ID NO: 4. In some embodiments, an antigenic portion of a Plasmodium TRAMP polypeptide comprises a glutamine at position 149, position 195, and position 202, as numbered according to SEQ ID NO: 4. In some embodiments, an antigenic portion of a Plasmodium TRAMP polypeptide comprises or consists of an amino acid sequence according to SEQ ID NO: 181.

[0475] 42-352 portion of TRAMP Aolvcan (SEQ ID NO: 181)

LQVYQNNGNFKMEILEPECnQKNLPSEEEKEEnDFDRKELLLHDFWIKNSQFTDEKTLELYLSKEENNKSYLMLTFYLGDLKLMIG HNSPYEISLILNVATVNKQMSNCKNNSYNIVLLIGTDVFNTSDLEILEGPIQFSLGKSSGAFRIQVTNFFLQNTWKAFTKDKISFLIKPE ENCYnLENKINKPQLUEKKTTFYSEWGEWSNCSMDCDHPDNVQIRERECIHPSGDCFKGDLKESRPCIIPLPPCNELFSHKDNSTF KILMIILPIVLVISimLYHIFYKRKGAEKELYENVAGRYMYD

[0476] In some embodiments, an antigenic portion of a Plasmodium TRAMP polypeptide comprises an asparagine at position 112, position 149, position 155, position 170, position 195, position 202, position 253, position 305, or a combination thereof, as numbered according to SEQ ID NO: 4. In some embodiments, an antigenic portion of a Plasmodium TRAMP polypeptide comprises a glutamine at position 112, position 149, position 155, position 170, position 195, position 202, position 253, position 305, or a combination thereof, as numbered according to SEQ ID NO: 4. In some embodiments, an antigenic portion of a Plasmodium TRAMP polypeptide comprises a glutamine at position 112, position 149, position 155, position 170, position 195, position 202, position 253, and position 305, as numbered according to SEQ ID NO: 4. In some embodiments, an antigenic portion of a Plasmodium TRAMP polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 180.

[0477] 42-352 portion of TRAMP Aolvcen (SEQ ID NO: 180)

LQVYQNNGNFKMEILEPECnQKNLPSEEEKEEFIDFDRKELLLHDFVVIKNSQFTDEKTLELYLSKEENQKSYLMLTFYLGDLKLMIG HNSPYEISLILNVATVNKQMSNCKQNSYNIVLLJGTDVFQTSDLEILEGPIQFSLGKSSGAFRIQVrNFFLQNTWKAFTKDKISFLIKPE ENCYTILENKINKPQLLIEKICTTFYSEWGEWSQCSMDCDHPDNVQIRERECIHPSGDCFKGDLKESRPCIIPLPPCNELFSHKDQSTF KUMHLPIVLVISimLYHIFYKRKGAEKELYENVAGRYMYD

[0478] In some embodiments, an antigenic portion of a Plasmodium TRAMP polypeptide comprises a PMX cleavage site. In some embodiments, a PMX cleavage site of TRAMP antigenic portion comprises or consists of a sequence of HFLQ (SEQ ID NO: 192). In some embodiments, an antigenic portion of a PlasmodkimlRMAP polypeptide comprises a SUB2 cleavage site. In some embodiments, a SUB2 cleavage site of TRAMP antigenic portion comprises or consists of ILMIILPIVLVISimLYHIFY (SEQ ID NO: 193).

[0479] In some embodiments, an antigenic portion of a Plasmodium TRAMP polypeptide comprises a transmembrane region (also referred to as a "transmembrane domain”). In some embodiments, an antigenic portion of a PiasmodiumTWM? polypeptide comprises a transmembrane region at the C-terminus. In some embodiments, an antigenic portion of a Plasmodium TRAMP polypeptide comprises a HSV transmembrane region, e.g., an HSV-1 or HSV-2 transmembrane region. In some embodiments, an antigenic portion of a Plasmodium TRAMP polypeptide comprises a transmembrane region that comprises or consists of an HSV gD transmembrane region, e.g., comprising or consisting of an amino add sequence of SEQ ID NO:75. In some embodiments, a Plasmodium TRAMP polypeptide does not comprise a transmembrane region.

6. CSS

[0480] In some embodiments, a Plasmodium polypeptide construct described herein Includes one or more Plasmodium Rh5 Invasion complex polypeptides or portions thereof (e.g., antigenic portions thereof) that comprises one or more Plasmodium cysteine-rich small secreted protein (CSS) polypeptides or antigenic portions thereof. [0481] In some embodiments, a Plasmodium polypeptide construct described herein includes one or more regions or portions of a CSS, e.g., Plasmodium CSS, e.g., P. falciparum CSS (SEQ ID NO: 5 or 214), or a variant thereof (e.g., one or more antigenic portions of a CSS, e.g., Plasmodium CSS, e.g., P. falciparum CSS").

[0482] In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide comprises a C30S mutation, as numbered according to SEQ ID NO: 5 or 214. In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide comprises a serine at position 30, as numbered according to SEQ ID NO: 5 or 214. [0483] In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide comprises or consists of an amino acid sequence with at least 85% sequence Identity to an amino acid sequence according to SEQ ID NO: 194. In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 194. In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 195. In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 195. In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 182. In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 182.

[0484] 21-152 portion of CSS (SEQ ID NO: 194)

QDEKSVKNISVCDfTDKLNFLPLEKTKILCELKPQYGEDIKIIANKEYEINCMNNSKVFSPLKDTFINMTNIKLYSPKLHFEIKDITHKGK NAALYYLKIDEEASDIFFSCSIKPKQVSGLLEGEVRVNLKK

[0485] 153-290 portion of CSS (SEQ ID NO: 195)

HINEEYSIFNEEEDVHVCDFSKGNLDUPSAGFYLKNSRNVSCIYRVIPNKLFUKLPKLDIVTEKLLPSIVNCLSEFSHNFTLKHVQEG DNYISFNVIFGEFKKHFNLTCSLDLSDFQQEPSNLGKTANITFIFSK

[0486] 21-290 portion of CSS (SEQ ID NO: 182)

QDEKSVKNISVCDFTDKLNFLPLEKTKILCELKPQYGEDIKIIANKEYEINCMNNSKVFCPLKDTFINNTNIKLYSPKLHFEIKDITHKGK NAALYYLKIDEEASDIFFSCSIKPKQVSGLLEGEVRVNLKKHINEEYSIFNEEEDVHVCDFSKGNLDITPSAGFYLKNSRNVSCIYRVIP NKLFLIKLPKLDIVTEKLLPSIVNCLSEFSnNFTLKHVQEGDNYISFNVIFGEFKKHFNLTCSLDLSDFQQEPCNLGKTANnrHFSK [0487] In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide comprises 25, 30, 35, 40, or 45 contiguous amino acids of a CSS polypeptide, or a variant thereof (e.g., a glycosylation variant). In some embodiments, a Plasmodium polypeptide construct comprises one or more antigenic portions of CSS.

[0488] In some embodiments, a Plasmodium CSS antigenic portion comprises one or more N-linked glycosylation sites. In some embodiments, a Plasmodium CSS antigenic portion comprises one, two, three, four, five, or six N-linked glycosylation sites. In some embodiments, a Plasmodium CSS antigenic portion comprises an amino add substitution at one or more N-llnked glycosylation sites, where the amino add substitution prevents glycosylation. In some embodiments, an amino acid substitution that prevents glycosylation comprises a NX[T/S] to QX[T/S] substitution. In some embodiments, amino acid substitution prevents glycosylation comprises a NX[T/S] to NXA substitution. In some embodiments, a Plasmodium CSS antigenic portion comprises an amino add substitution at all of the N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation.

[0489] In some embodiments, an antigenic portion of Plasmodium CSS polypeptide comprises a serine at position 80, as numbered according to SEQ ID NO: 5 or 214.

[0490] In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide comprises an asparagine at position 74, position 88, or a combination thereof, as numbered according to SEQ ID NO: 5 or 214. In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide comprises a glutamine at position 74, position 88, or a combination thereof, as numbered according to SEQ ID NO: 5 or 214. In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide comprises a glutamine at position 74, and position 88, as numbered according to SEQ ID NO: 5 or 214. In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide comprises or consists of an amino acid sequence according to SEQ ID NO: 184.

[0491] 21-152 portion of CSS Advcan (SEQ ID NO: 184)

QDEKSVKNISVCDFTDKLNFLPLEKTKILCELKPQYGEDIKIIANKEYBNCMQNSKVFSPLKDTFIQNTNIKLYSPKI-HFEIKDITHKGK NAALYYLKIDEEASDIFFSCSIKPKQVSGLLEGEVRVNLKK

[0492] In some embodiments, an antigenic portion of Plasmodium CSS polypeptide comprises a serine at position 276, as numbered according to SEQ ID NO: 5 or 214. In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide comprises an asparagine at position 192, position 234, position 261, position 283, or a combination thereof, as numbered according to SEQ ID NO: 5 or 214. In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide comprises a glutamine at position 192, position 234, position 261, position 283, or a combination thereof, as numbered according to SEQ ID NO: 5 or 214. In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide comprises a glutamine at position 192, position 234, position 261, and position 283, as numbered according to SEQ ID NO: 5 or 214. In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide comprises or consists of an amino acid sequence according to SEQ ID NO: 185.

[0493] 153-290 portion of CSS Aolvcan (SEQ ID NO: 185)

HINEEYSIFNEEEDVHVCDFSKGNLDITPSAGFYLKNSRQVSCIYRVIPNKLFUKLPKLDIVTEKLLPSIVNCLSEFSHQFTLKHVQEG DNYISFNVIFGEFKKHFQLTCSLDLSDFQQEPSNLGKrAQITFIFSK

[0494] In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide comprises an asparagine at position 74, position 88, position 192, position 234, position 261, position 283, or a combination thereof, as numbered according to SEQ ID NO: 5 or 214. In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide comprises a glutamine at position 74, position 88, position 192, position 234, position 261, position 283, or a combination thereof, as numbered according to SEQ ID NO: 5 or 214. In some embedments, an antigenic portion of a Plasmodium CSS polypeptide comprises a glutamine at position 74, position 88, position 192, position 234, position 261, and position 283, as numbered according to SEQ ID NO: 5 or 214. In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 183.

[0495] 21-290 portion of CSS Aolvcan (SEQ ID NO: 183)

QDEKSVKNISVCDFTDKLNFLPLEKTKILCELKPQYGEDIKIIANKEYEINCMQNSKVFCPLKDTFJQNTNIKLYSPKLHFBKDITHKGK NMLYYLKIDEEASDIFFSCSIKPKQVSGLLEGEVRVNLKKHINEEYSIFNEEEDVHVCDFSKGNLDnPSAGFYLKNSRQVSCIYRVIP NKLFLIKLPKLDIVTEKLLPSIVNCLSEFSFIQFTLKHVQEGDNYISFNVIFGEFKKHFQLTCSLDLSDFQQEPCNLGKTAQITIFSK B. Secretory Signals

[0496] In some embodiments, a Plasmodium polypeptide construct described herein Includes a secretory signal, e.g., that Is functional In mammalian cells. In some embodiments, a utilized secretory signal is a heterologous secretory signal. In some embodiments, a heterologous secretory signal comprises or consists of a non-human secretory signal. In some embodiments, a heterologous secretory signal comprises or consists of a viral secretory slgnal. In some embodiments, a viral secretory signal comprises or consists of an hSV secretory signal (e.g., an HSV- 1 or HSV-2 secretory signal). In some embodiments, an HSV secretory signal comprises or consists of an HSV glycoprotein D (gD) secretory signal.

[0497] In some embodiments, a seaetory signal comprises or consists of an Ebola virus secretory signal. In some embodiments, an Ebola virus secretory signal comprises or consists of an Ebola virus spike glycoprotein (SGP) secretory signal.

[0498] In some embodiments, a secretory signal Is characterized by a length of about 15 to 30 amino adds. [0499] In many embodiments, a secretory signal is positioned at the N-terminus of a Plasmodium polypeptide construct described herein. In some embodiments, a secretory signal preferably allows transport of a Plasmodium polypeptide construct with which it is assodated into a defined cellular compartment, preferably a cell surface, endoplasmic reticulum (ER) or endosomal-lysosomal compartment [0500] In some embodiments, a secretory signal comprises or consists of a Plasmodium secretory signal. In some embodiments, a Plasmodium seaetory signal comprises or consists of a Plasmodium CSP secretory signal (e.g., from Plasmodium fafdpanim, e.g., from Plasmodium falciparumVsfitete 3D7 (SEQ ID NO: 7)). In some embodiments, a Plasmodium seaetory signal comprises a Plasmodium Rh5 seaetory signal. In some embodiments, a Plasmodium secretory signal comprises a Plasmodium CyRPA secretory signal. In some embodiments, a Plasmodium sec.xe.torf signal comprises or consists of an amino acid sequence according to SEQ ID NO: 187. In some embodiments, a Piasmocfium secretory signal comprises a Plasmodium Pl 13 secretory signal. In some embodiments, a Plasmodium secretory signal comprises a Plasmodium Ripr secretory signal. In some embodiments, a Plasmodium secretory signal comprises or consists of an amino add sequence according to SEQ ID NO: 186. In some embodiments, a Plasmodium secretory signal comprises a Piasmocfium TRAMP secretory signal. In some embodiments, a Plasmodium secretory signal comprises a Plasmodium CSS secretory signal. In some embodiments, a Plasmodium secretory signal comprises or consists of an amino acid sequence according to SEQ ID NO: 188.

[0501] In some embodiments, a seaetory signal is selected from an S1S2 secretory signal (aa 1-19), an Immunoglobulin secretory signal (aa 1-22), a human SPARC secretory signal, a human Insulin Isoform 1 seaetory signal, a human albumin secretory signal, etc. Those skilled in the art will be aware of other secretory signal such as, for example, as disclosed in W02017/081082, which is incorporated herein by reference in its entirety (e.g., SEQ ID NOs: 1-1115 and 1728, or fragments variants thereof). In some embodiments, a Plasmodium polypeptide construct described herein does not comprise a secretory signal.

[0502] In some embodiments, a seaetory signal is one listed in Table 3, or a secretory signal having 1, 2, 3, 4, or 5 amino acid differences relative thereto. In some embodiments, a signal sequence is selected from those Included In the Table 3 below and/or those encoded by the sequences In Table 4 below.

Table 3: Exemplary secretory signals Table 4: Exemplary polynucleotide sequences encoding secretory signals

C. Transmembrane Regions

[0503] In some embodiments, a Plasmodium polypeptide construct described herein Includes a transmembrane region (also referred to as a "transmembrane domain”). In some embodiments, a transmembrane region is located at the N-termlnus of a Plasmodium polypeptide construct In some embodiments, a transmembrane region Is located at the C-terminus of a Plasmodium polypeptide construct. In some embodiments, a transmembrane region is not located at the N-terminus or C-terminus of a Plasmodium polypeptide construct.

[0504] Transmembrane regions are known in the art, any of which can be utilized in a Plasmodium polypeptide construct described herein. In some embodiments, a transmembrane region comprises or is a transmembrane domain of Hemagglutinin (HA) of Influenza virus, Env of HIV-1, equine infectious anaemia virus (EIAV), murine leukaemia virus (MLV), mouse mammary tumor virus, G protein of vesicular stomatitis virus (VSV), Rabies virus, or a seven transmembrane domain receptor.

[0505] In some embodiments, a heterologous transmembrane region does not comprise a hemagglutin transmembrane region. In some embodiments, a heterologous transmembrane region comprises or consists of a non-human transmembrane region. In some embodiments, a heterologous transmembrane region comprises or consists of a viral transmembrane region. In some embodiments, a heterologous transmembrane region comprises or consists of an HSV transmembrane region, e.g., an HSV-1 or HSV-2 transmembrane region. In some embodiments, an HSV transmembrane region comprises or consists of an HSV gD transmembrane region, e.g., comprising or consisting of an amino add sequence of GLIAGAVGGSLLAALVICGIVYWMRRHTQKAPKRIRLPHIR (SEQ ID NO: 75). [0506] In some embodiments, a heterologous transmembrane region comprises or consists of a human transmembrane region. In some embodiments, a human transmembrane region comprises or consists of a human decay accelerating factor glycosylphosphatidylinositol (hDAF-GPI) anchor region. In some embodiments, an hDAF- GPI anchor region comprises or consists of an amino add sequence of PNKGSGTTSGTTRLi-SGHTCFTLTGLLGTLVTMGLLT (SEQ ID NO:76).

[0507] In some embodiments, a transmembrane region comprises or consists of a Plasmodium transmembrane region. In some embodiments, a utilized transmembrane region Is one that is normally associated with CSP In nature. In some embodiments, a Plasmodium transmembrane region comprises or consists of a Plasmodium QSP glycosylphosphatidylinositol (GPI) anchor region. In some embodiments, a Plasmodium CSP GPI anchor region is from Plasmodium falciparum. In some embodiments, a Plasmodium CSP GPI anchor region is from Plasmodium falciparum isolate 3D7 (SEQ ID N0:7), e.g., amino adds 374-397 of SEQ ID N0:7.

[0508] In some embodiments, a transmembrane region comprises PlasmodiumTRAP transmembrane region. In some embodiments, a transmembrane region comprises Plasmodium P113 transmembrane region.

[0509] In some embodiments, a utilized transmembrane region is a heterologous transmembrane region. [0510] In some embodiments, a Plasmodium polypeptide construct described herein does not comprise a transmembrane region.

[0511] Exemplary transmembrane are provided In the following Table 5: Table 5: Exemplary transmembrane regions

D. Multimerization Regions

[0512] In some embodiments, a Plasmodium polypeptide construct described herein Includes one or more multimerization regions (e.g., a heterologous multimerization region). In some embodiments, a heterologous multimerization region comprises a dimerization, trimerizatlon or tetramerization region.

[0513] In some embodiments, a multimerization region is one described in W02017/081082, which is Incorporated herein by reference In Its entirety (e.g., SEQ ID NOs: 1116-1167, or fragments or variants thereof). Exemplary trimerizatlon and tetramerization regions Include, but are not limited to, engineered leucine zippers, fibritki fbldon domain from enterobacteria phage T4, GCN4pll, GCN4-pll, and p53.

[0514] In some embodiments, a provided Plasmodium polypeptide construct described herein Is able to form a trimeric complex. For example, a provided Plasmodium polypeptide construct may comprise a multimerization region allowing formation of a multimeric complex, such as for example a trimeric complex of a Plasmodium polypeptide construct described herein. In some embodiments, a multimerization region allowing formation of a multimeric complex comprises a trimerization region, for example, a trimerizatlon region described herein. In some embodiments, a Plasmodium polypeptide construct includes a T4-fibritin-derived '"fbldon"' trimerization region, for example, to increase its immunogenicity. In some embodiments, a Plasmodium polypeptide construct includes a multimerization region comprising or consisting of the amino acid sequence GYIPEAPRDGQAYVRKDGEWVLLSTFL (SEQ ID NO: 78).

E. Linkers

[0515] In some embodiments, a Plasmodium polypeptide construct described herein includes one or more linkers. In some embodiments, a linker is or comprises 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acids. In some embodiments, a linker is or comprises no more than about 30, 25, 20, 15, 10 or fewer amino adds. A linker can indude any amino acid sequence and is not limited to any particular amino adds. In some embodiments, a linker comprises one or more gtydne (G) amino acids. In some embodiments, a linker comprises one or more serine (S) amino acids. In some embodiments, a linker comprises a glycine-serlne linker. A “glydne-serlne inker" as used herein refers to a linker that comprises predominantly (e.g., 80% or more) glyche end serine amino acids. In some embodiments, a linker indudes amino adds selected based on a deavage predictor to generate highly-deavable linkers.

[0516] In some embodiments, a linker is or comprises S-G4-S-G4-S. In some embodiments, a linker Is or comprises GSPGSGSGS (SEQ ID NO: 79). In some embodiments, a linker is or comprises GGSGGGGSGG (SEQ ID NO: 80). In some embodiments, a linker Is one presented in Table 6. In some embodiments, a linker is or comprises a sequence as set forth in W02017/081082, which Is incorporated herein by reference In Its entirety (see SEQ ID NOs: 1509-1565, or a fragment or variant thereof).

[0517] In some embodiments, a Plasmodium polypeptide construct described herein comprises a linker between a C-terminal region or portion thereof and a transmembrane region. In some embodiments, a Plasmodium polypeptide construct described herein comprises a linker after a minor repeat sequence.

[0518] Exemplary linkers are provided In the following Table 6:

Table 6: Exemplary linkers

SEQ ID NO: Sequence (Amino Add)

81 SGGGGSGGGGS

79 GSPGSGSGS

80 GGSGGGGSGG

82 GGS

83 GGGS

84 GGGGSGGGGSGGGGS

85 AGNRVRRSVG

86 GSGSGS

87 GGSLGGGGSG

137 SGG

F. Setf-Assembling Regions

[0519] In some embodiments, a Plasmodium polypeptide construct described herein Includes one or more selfassembling regions (e.g., a self-assembling nanopartide region, e.g., a heterologous self-assembling nanoparticle region). In some embodiments, a self-assembling nanoparticle region is a ferritin region. In some embodiments, a ferritin region is from H. pylori. In some embodiments, a ferritin region comprises or consists of a sequence of: DIIKLLNEQVNKEMQSSNLYMSMSSWCYTHSLDGAGLFLFDHAAEEYEHAKKUIFLNENNVPVQLTSISAPEHKFEGLTQIFQKAYE HEQHISESINNIVDHAIKSKDHATFNFLQWYVAEQHEEEVLFKDILDKIEUGNENHGLYLADQYVKGIAKSRKS (SEQ ID NO: 88). G. Tags

[0520] Those skilled in the art, reading the present disclosure, will appreciate that, in some embodiments, one or more tags may be used when designing and testing constructs (e.g., Plasmodium polypeptide constructs described herein) in certain contexts (e.g., in vitro, ex vivo, etc.). In some embodiments, one or more tags may be directly connected to a Plasmodium polypeptide construct through a peptide bond (e.g., at the S'-end or 3'-end of a construct). In some embodiments, one or more tags may be connected to a Plasmodium polypeptide construct through one or more linkers (e.g., one or more linkers described herein). In some embodiments, one or more tags may be internally embedded within a Plasmodium polypeptide construct, wherein the one or more tags are directly connected to the construct through peptide bonds (e.g., at the S'-end and 3'-end of the one or more tags). In some embodiments, one or more tags may be Internally embedded within a Plasmodium polypeptide construct, wherein the one or more tags are connected to the construct through one or more linkers (e.g., one or more linkers described herein).

[0521] In some embedments, a Plasmodium polypeptide construct described herein Includes one or more tags. In some embodiments, a Plasmodium polypeptide construct described herein Includes one or more detection tags (e.g., HiBit tag, HA tag, etc.). In some embodiments, a Plasmodium polypeptide construct described herein indudes a tag that comprises or consists of a HiBit tag. In some embodiments, a HiBit tag has an amino add sequence according to SEQ ID NO: 139.

H. Embodiments of Plasmodium polypeptide constructs

[0522] In some embodiments, a Plasmodium polypeptide construct described herein includes one or more

Plasmodium Rh5 Invasion complex polypeptide regions or portions thereof as described above. Exemplary combinations of regions are described below.

[0523] In some embodiments, a Plasmodium polypeptide construct described herein Includes one or more regions or portions of a Rh5 invasion complex polypeptide from Plasmodium fatoparum, preferably from Plasmodium fatoparum isolate 3D7. In some embodiments, a Rh5 Invasion complex polypeptide is selected from: Plasmodium Rh5, Plasmodium CyRPA, Plasmodium Ripr, Plasmodium P113, Plasmodium TRAMP, and Plasmodium CSS.

1. Rh5 constructs

[0524] In some embodiments, the Plasmodium polypeptide construct described herein Includes an ordered domain of Rh5 from Plasmodium fatoparum, preferably from Plasmodium faidparum isolate 3D7. In some embodiments, the Plasmodium polypeptide construct described herein includes an ordered domain of Rh5 from Plasmodium faidparum, preferably from Plasmodium fatoparum isolate 3D7 (referred to herein as a "Rh5 construct" or "Rh5 ordered domain").

[0525] In some embodiments, an ordered domain of Rh5 comprises the amino add sequence of positions 140- 247 of SEQ ID NO: 1, or an amino add sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the amino add sequence of positions 140-247 of SEQ ID NO: 1. In some embodiments, an ordered domain of Rh5 comprises the amino add sequence of positions 140-247 of SEQ ID NO: 1, or the amino add sequence of positions 140-247 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino acid substitutions.

[0526] In some embodiments, an ordered domain of Rh5 comprises the amino acid sequence of positions 146- 247 of SEQ ID NO: 1, or an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the amino add sequence of positions 146-247 of SEQ ID NO: 1. In some embodiments, an ordered domain of Rh5 comprises the amino add sequence of positions 146-247 of SEQ ID NO: 1, or the amino acid sequence of positions 146-247 of SEQ ID NO:1 having 1, 2, 3, 4, or 5 amino acid substitutions.

[0527] In some embodiments, an ordered domain of Rh5 comprises the amino acid sequence of positions 297- 526 of SEQ ID NO: 1, or an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the amino add sequence of positions 297-526 of SEQ ID NO: 1. In some embodiments, an ordered domain of Rh5 comprises the amino add sequence of positions 297-526 of SEQ ID NO: 1, or the amino add sequence of positions 297-526 of SEQ ID NO:1 having 1, 2, 3, 4, or 5 amino acid substitutions.

[0528] In some embodiments, an ordered domain of Rh5 can have the following structure: sequence corresponding to amino adds 140-247 of SEQ ID NO: 1 (140-247 Ordered Domain); sequence corresponding to amino acids 140-247 of SEQ ID NO: 1 with a tyrosine at position 203 (140- 247 Ordered Domain-203Y); sequence corresponding to amino acids 140-247 of SEQ ID NO: 1 with a substitution that prevents glycosylation at position 214 (140-247 Ordered Domain-A214gly); sequence corresponding to amino acids 140-247 of SEQ ID NO: 1 with a tyrosine at position 203 and a substitution that prevents glycosylation at position 214 (140-247 Ordered Domain-203Y-A214gly); sequence corresponding to amino acids 140-247 of SEQ ID NO: 1 with a glutamine at position 214 (140-247 Ordered Domaln-N214Q); sequence corresponding to amino acids 140-247 of SEQ ID NO: 1 with a tyrosine at position 203 and a glutamine at position 214 (140-247 Ordered Domain-203Y-N214Q); sequence corresponding to amino adds 140-247 of SEQ ID NO: 1 with a substitution that prevents PMX deavage (140-247 Ordered Domaln-APMX); sequence corresponding to amino acids 140-247 of SEQ ID NO: 1 with a substitution that prevents PMX deavage and a tyrosine at position 203 and a glutamine at position 214 (140-247 Ordered Domaki-APMX-203Y- N214Q); sequence corresponding to amino adds 146-247 of SEQ ID NO: 1 (146-247 Ordered Domain); sequence corresponding to amino acids 146-247 of SEQ ID NO: 1 with a tyrosine at position 203 (146- 247 Ordered Domaln-203Y); sequence corresponding to amino adds 146-247 of SEQ ID NO: 1 with a substitution that prevents glycosylation at position 214 (146-247 Ordered Domain-A214gly); sequence corresponding to amino acids 146-247 of SEQ ID NO: 1 with a tyrosine at position 203 and a substitution that prevents glycosylation at position 214 (146-247 Ordered Domain-203Y-A214gly); sequence corresponding to amino adds 146-247 of SEQ ID NO: 1 with a glutamine at position 214 (146-247 Ordered Domain-N214Q); sequence corresponding to amino adds 146-247 of SEQ ID NO: 1 with a tyrosine at position 203 and a glutamine at position 214 (146-247 Ordered Domain-203Y-N214Q); sequence corresponding to amino adds 297-526 of SEQ ID NO: 1 (297-526 Ordered Domain); sequence corresponding to amino adds 297-526 of SEQ ID NO: 1 with a tyrosine at position 329 (297- 526 Ordered Domain-329Y); sequence corresponding to amino adds 297-526 of SEQ ID NO: 1 with a substitution that prevents glycosylation at position 297 (297-526 Ordered Domain-A297gly); sequence corresponding to amino acids 297-526 of SEQ ID NO: 1 with a tyrosine at position 329 and a substitution that prevents glycosylation at position 297 (297-526 Ordered Domain-329Y-A297gly); sequence corresponding to amino acids 297-526 of SEQ ID NO: 1 with a glutamine at position 297 (297-526 Ordered Domain-N297Q); or sequence corresponding to amino acids 297-526 of SEQ ID NO: 1 with a tyrosine at position 329 and a glutamine at position 297 (297-526 Ordered Domain- 329Y-N297Q).

[0529] In some embodiments, a Plasmodium polypeptide construct described herein includes two ordered domains of Rh5. In some embodiments, two ordered domains of Rh5 are from Plasmodium fakjparum, preferably from Plasmodium fakjparum isolate 3D7.

[0530] In some embodiments, a Plasmodium polypeptide construct comprises a Rh5 polypeptide or antigenic portion thereof that includes 2 ordered domains of Rh5. In some embodiments, an Rh5 portion of a malarial polypeptide construct includes two ordered domains of Rh5 that have the following structure:

Rh5 ordered domain Constructs

140-247 Ordered Domain - 297-526 Ordered Domain;

140-247 Ordered Domain-203Y - 297-526 Ordered Domain;

140-247 Ordered Domain-A214gly - 297-526 Ordered Domain;

140-247 Ordered Domain-203Y-A214gly - 297-526 Ordered Domain;

140-247 Ordered DomainAPMX - 297-526 Ordered Domain;

140-247 Ordered DomainAPMX-203Y - 297-526 Ordered Domain;

140-247 Ordered DomainAPMX-A214gly - 297-526 Ordered Domain;

140-247 Ordered DomainAPMX-203Y-A214gly - 297-526 Ordered Domain;

146-247 Ordered Domain - 297-526 Ordered Domain;

146-247 Ordered Domain-203Y - 297-526 Ordered Domain;

146-247 Ordered Domaln-A214gly - 297-526 Ordered Domain;

146-247 Ordered Domain-203Y-A214gly - 297-526 Ordered Domain;

140-247 Ordered Domain - 297-526 Ordered Domain-329Y;

140-247 Ordered Domain-203Y - 297-526 Ordered Domain-329Y;

140-247 Ordered Domain-A214gly - 297-526 Ordered Domain-329Y;

140-247 Ordered Domain-203Y-A214gly - 297-526 Ordered Domain-329Y;

140-247 Ordered DomainAPMX - 297-526 Ordered Domain-329Y;

140-247 Ordered DomainAPMX-203Y - 297-526 Ordered Domain-329Y;

140-247 Ordered DomalnAPMX-A214gly - 297-526 Ordered Domain-329Y;

140-247 Ordered DomainAPMX-203Y-A214gly - 297-526 Ordered Domain-329Y;

146-247 Ordered Domain - 297-526 Ordered Domain-329Y;

146-247 Ordered Domain-203Y - 297-526 Ordered Domain-329Y;

146-247 Ordered Domain-A214gly - 297-526 Ordered Domain-329Y;

146-247 Ordered Domain-203Y-A214gly - 297-526 Ordered Domain-329Y;

140-247 Ordered Domain - 297-526 Ordered Domain A297gly;

140-247 Ordered Domain-203Y - 297-526 Ordered DomainA297gly;

140-247 Ordered Domain-A214gly - 297-526 Ordered Domain A297gly; 140-247 Ordered Domain-203Y-A214gly - 297-526 Ordered Domain A297gly;

140-247 Ordered DomainAPMX - 297-526 Ordered Domain A297gly;

140-247 Ordered DomainAPMX-203Y - 297-526 Ordered Domain A297gly;

140-247 Ordered DomainAPMX-A214gly - 297-526 Ordered Domain A297gly;

140-247 Ordered DomainAPMX-203Y-A214gly - 297-526 Ordered Domain A297gly;

146-247 Ordered Domain - 297-526 Ordered Domain A297gly;

146-247 Ordered Domain-203Y - 297-526 Ordered Domain A297gly;

146-247 Ordered Domain-A214gly - 297-526 Ordered Domain A297gly;

146-247 Ordered Domain-203Y-A214gly - 297-526 Ordered DomainA297gly;

140-247 Ordered Domain - 297-526 Ordered Domain-329YA297gly;

140-247 Ordered Domain-203Y - 297-526 Ordered Domain-329YA297gly;

140-247 Ordered Domain-A214gly - 297-526 Ordered Domain-329YA297gly;

140-247 Ordered Domain-203Y-A214gly - 297-526 Ordered Domain-329YA297gly;

140-247 Ordered DomainAPMX - 297-526 Ordered Domain-329YA297gly;

140-247 Ordered DomainAPMX-203Y - 297-526 Ordered Domain-329YA297gly;

140-247 Ordered DomainAPMX-A214gly - 297-526 Ordered Domain-329YA297gly;

140-247 Ordered DomalnAPMX-203Y-A214gly - 297-526 Ordered Domaln-329YA297gly;

146-247 Ordered Domain - 297-526 Ordered Domaln-329YA297gly;

146-247 Ordered Domain-203Y - 297-526 Ordered Domaln-329YA297gly;

146-247 Ordered Domain-A214gly - 297-526 Ordered Domain-329YA297gly; or

146-247 Ordered Domain-203Y-A214gly - 297-526 Ordered Domain-329YA297gly.

[0531] In some embodiments, Plasmodium polypeptide construct indudes an Rh5 polypeptide or antigenic portion thereof that Indudes 2 ordered domains of Rh5 and a linker. In some embodiments, an Rh5 portion of a maliarlal polypeptide has the following structure:

RhS ordered domain with Linker Constructs

140-247 Ordered Domain - linker - 297-526 Ordered Domain;

140-247 Ordered Domain-203Y - linker - 297-526 Ordered Domain;

140-247 Ordered Domain-A214gly - linker - 297-526 Ordered Domain;

140-247 Ordered Domain-203Y-A214gly - linker - 297-526 Ordered Domain;

140-247 Ordered DomainAPMX - linker - 297-526 Ordered Domain;

140-247 Ordered DomainAPMX-203Y - linker - 297-526 Ordered Domain;

140-247 Ordered DomainAPMX-A214gly - linker - 297-526 Ordered Domain;

140-247 Ordered DomainAPMX-203Y-A214gly - linker - 297-526 Ordered Domain;

146-247 Ordered Domain - linker - 297-526 Ordered Domain;

146-247 Ordered Domain-203Y - linker - 297-526 Ordered Domain;

146-247 Ordered Domain-A214gly - linker - 297-526 Ordered Domain;

146-247 Ordered Domain-203Y-A214gly - linker - 297-526 Ordered Domain;

140-247 Ordered Domain - linker - 297-526 Ordered Domain-329Y;

140-247 Ordered Domain-203Y - linker - 297-526 Ordered Domain-329Y;

140-247 Ordered Domain-A214gly - linker - 297-526 Ordered Domain-329Y; 140-247 Ordered Domain-203Y-A214gly - linker - 297-526 Ordered Domain-329Y;

140-247 Ordered DomainAPMX - linker - 297-526 Ordered Domain-329Y;

140-247 Ordered DomainAPMX-203Y - linker - 297-526 Ordered Domain-329Y;

140-247 Ordered DomainAPMX-A214gly - linker - 297-526 Ordered Domain-329Y;

140-247 Ordered DomainAPMX-203Y-A214gly - linker - 297-526 Ordered Domain-329Y;

146-247 Ordered Domain - linker - 297-526 Ordered Domain-329Y;

146-247 Ordered Domain-203Y - linker • 297-526 Ordered Domain-329Y;

146-247 Ordered Domain-A214gly - linker - 297-526 Ordered Domain-329Y;

146-247 Ordered Domain-203Y-A214gly - linker - 297-526 Ordered Domaln-329Y;

140-247 Ordered Domain - linker - 297-526 Ordered Domain A297gly;

140-247 Ordered Domain-203Y - linker - 297-526 Ordered DomainA297gty;

140-247 Ordered Domain-A214gly - linker - 297-526 Ordered Domain A297gly;

140-247 Ordered Domain-203Y-A214gly - linker - 297-526 Ordered Domain A297gly;

140-247 Ordered DomainAPMX - linker - 297-526 Ordered Domain A297gly;

140-247 Ordered DomainAPMX-203Y - linker - 297-526 Ordered Domain A297gly;

140-247 Ordered DomainAPMX-A214gly - linker - 297-526 Ordered Domain A297gly;

140-247 Ordered DomalnAPMX-203Y-A214gly - linker - 297-526 Ordered Domain A297gly;

146-247 Ordered Domain - linker - 297-526 Ordered Domain A297gly;

146-247 Ordered Domain-203Y - linker - 297-526 Ordered Domain A297gly;

146-247 Ordered Domaln-A214gly - linker - 297-526 Ordered Domain A297gly;

146-247 Ordered Domain-203Y-A214gly - linker - 297-526 Ordered DomalnA297gly;

140-247 Ordered Domain - linker - 297-526 Ordered Domain-329YA297gly;

140-247 Ordered Domain-203Y - linker - 297-526 Ordered Domain-329YA297gly;

140-247 Ordered Domaln-A214gly - linker - 297-526 Ordered Domaln-329YA297gly;

140-247 Ordered Domain-203Y-A214gly - linker - 297-526 Ordered Domain-329YA297gly;

140-247 Ordered DomainAPMX - linker - 297-526 Ordered Domaln-329YA297gly;

140-247 Ordered DomainAPMX-203Y - linker - 297-526 Ordered Domain-329YA297gly;

140-247 Ordered DomainAPMX-A214gly - linker - 297-526 Ordered Domain-329YA297gly;

140-247 Ordered DomainAPMX-203Y-A214gly - linker - 297-526 Ordered Domain-329YA297gly;

146-247 Ordered Domain - linker - 297-526 Ordered Domain-329YA297gly;

146-247 Ordered Domain-203Y - linker - 297-526 Ordered Domain-329YA297gly;

146-247 Ordered Domaln-A214gly - linker - 297-526 Ordered Domain-329YA297gly; or 146-247 Ordered Domain-203Y-A214gly - linker - 297-526 Ordered Domain-329YA297gly.

[0532] In some embodiments, Plasmodium polypeptide construct includes an Rh5 polypeptide or antigenic portions thereof that includes 2 ordered domains of Rh5 and one or more disordered domains. In some embodiments, a disordered domain is an L-disordered domain of Rh5. In some embodiments, an L-disordered domain comprises a sequence corresponding to amino acids 248-296 of SEQ ID NO: 1. In some embodiments, a disordered domain is an L-disordered domain of Rh5. In some embodiments, an N-disordered domain comprises a sequence corresponding to amino adds 26-139 of SEQ ID NO: 1. In some embodiments, an Rh5 portion comprises both an L-disordered domain and an N-disordered domain of Rh5. [0533] In some embodiments, an Rh5 portion of a malarial polypeptide has two ordered domains and one or two disordered domains of Rh5. In some embodiments, an Rh5 portion of a malarial polypeptide has the following structure:

RhS ordered domains with L-Disordered Domain Constructs

140-247 Ordered Domain • L-Disordered Domain - 297-526 Ordered Domain;

140-247 Ordered Domain-203Y - L-Disordered Domain • 297-526 Ordered Domain;

140-247 Ordered Domain-A214gly - L-Disordered Domain - 297-526 Ordered Domain;

140-247 Ordered Domain-203Y-A214gly - L-Disordered Domain - 297-526 Ordered Domain;

140-247 Ordered DomainAPMX - L-Disordered Domain - 297-526 Ordered Domain;

140-247 Ordered DomalnAPMX-203Y - L-Disordered Domain - 297-526 Ordered Domain;

140-247 Ordered DomainAPMX-A214gly - L-Disordered Domain - 297-526 Ordered Domain;

140-247 Ordered DomainAPMX-203Y-A214gly - L-Disordered Domain - 297-526 Ordered Domain;

146-247 Ordered Domain - L-Disordered Domain - 297-526 Ordered Domain;

146-247 Ordered Domain-203Y - L-Disordered Domain - 297-526 Ordered Domain;

146-247 Ordered Domain-A214gly - L-Disordered Domain - 297-526 Ordered Domain;

146-247 Ordered Domain-203Y-A214gly - L-Disordered Domain - 297-526 Ordered Domain;

140-247 Ordered Domain - L-Disordered Domain - 297-526 Ordered Domaln-329Y;

140-247 Ordered Domain-203Y - L-Disordered Domain - 297-526 Ordered Domain-329Y;

140-247 Ordered Domain-A214gly - L-Disordered Domain - 297-526 Ordered Domaln-329Y;

140-247 Ordered Domain-203Y-A214gly - L-Disordered Domain - 297-526 Ordered Domaln-329Y;

140-247 Ordered DomainAPMX - L-Disordered Domain - 297-526 Ordered Domain-329Y;

140-247 Ordered DomalnAPMX-203Y - L-Disordered Domain - 297-526 Ordered Domain-329Y;

140-247 Ordered DomainAPMX-A214gly - L-Disordered Domain - 297-526 Ordered Domaln-329Y;

140-247 Ordered DomalnAPMX-203Y-A214gly - L-Dlsordered Domain - 297-526 Ordered Domaln-329Y;

146-247 Ordered Domain - L-Disordered Domain - 297-526 Ordered Domain-329Y;

146-247 Ordered Domain-203Y - L-Disordered Domain - 297-526 Ordered Domain-329Y;

146-247 Ordered Domain-A214gly - L-Disordered Domain - 297-526 Ordered Domain-329Y;

146-247 Ordered Domain-203Y-A214gly - L-Disordered Domain - 297-526 Ordered Domain-329Y;

140-247 Ordered Domain - L-Disordered Domain - 297-526 Ordered Domain A297gly;

140-247 Ordered Domain-203Y - L-Disordered Domain - 297-526 Ordered DomainA297gly;

140-247 Ordered Domain-A214gly - L-Disordered Domain - 297-526 Ordered Domain A297gly;

140-247 Ordered Domain-203Y-A214gly - L-Disordered Domain - 297-526 Ordered Domain A297gly;

140-247 Ordered DomainAPMX - L-Disordered Domain - 297-526 Ordered Domain A297gly;

140-247 Ordered DomainAPMX-203Y - L-Disordered Domain - 297-526 Ordered Domain A297gly;

140-247 Ordered DomainAPMX-A214gly - L-Disordered Domain - 297-526 Ordered Domain A297gly;

140-247 Ordered DomainAPMX-203Y-A214gly - L-Disordered Domain - 297-526 Ordered Domain A297gly;

146-247 Ordered Domain - L-Disordered Domain - 297-526 Ordered Domain A297gly;

146-247 Ordered Domain-203Y - L-Disordered Domain - 297-526 Ordered Domain A297gly;

146-247 Ordered Domain-A214gly - L-Disordered Domain - 297-526 Ordered Domain A297gly;

146-247 Ordered Domain-203Y-A214gly - L-Disordered Domain - 297-526 Ordered DomainA297gly; 140-247 Ordered Domain - L-Disordered Domain - 297-526 Ordered Domain-329YA297gly;

140-247 Ordered Domain-203Y - L-Disordered Domain - 297-526 Ordered Domain-329YA297gly;

140-247 Ordered Domain-A214gly - L-Disordered Domain - 297-526 Ordered Domain-329YA297gly;

140-247 Ordered Domain-203Y-A214gly • L-Disordered Domain - 297-526 Ordered Domain 329YA297gly;

140-247 Ordered DomainAPMX - L-Disordered Domain - 297-526 Ordered Domain-329YA297gly;

140-247 Ordered DomainAPMX-203Y - L-Disordered Domain - 297-526 Ordered Domain-329YA297gly;

140-247 Ordered DomainAPMX-A214gly - L-Disordered Domain - 297-526 Ordered Domain-329YA297gly;

140-247 Ordered DomainAPMX-203Y-A214gly - L-Disordered Domain - 297-526 Ordered Domain 329YA297gly;

146-247 Ordered Domain - L-Dlsordered Domain - 297-526 Ordered Domain-329YA297gly;

146-247 Ordered Domaln-203Y - L-Disordered Domain - 297-526 Ordered Domaln-329YA297gly;

146-247 Ordered Domain-A214gly - L-Disordered Domain - 297-526 Ordered Domain-329YA297gly; or

146-247 Ordered Domain-203Y-A214gly - L-Disordered Domain - 297-526 Ordered Domain-329YA297gly.

Rh5 ordered domains with N-Disordered Domain Constructs

N-Disordered Domain - 140-247 Ordered Domain - 297-526 Ordered Domain;

N-Disordered Domain - 140-247 Ordered Domain-203Y - 297-526 Ordered Domain;

N-Disordered Domain - 140-247 Ordered Domain-A214gly - 297-526 Ordered Domain;

N-Disordered Domain - 140-247 Ordered Domain-203 Y-A214gly - 297-526 Ordered Domain;

N-Disordered Domain - 140-247 Ordered DomainAPMX - 297-526 Ordered Domain;

N-Disordered Domain - 140-247 Ordered DomainAPMX-203Y - 297-526 Ordered Domain;

N-Disordered Domain - 140-247 Ordered DomalnAPMX-A214gly - 297-526 Ordered Domain;

N-Disordered Domain - 140-247 Ordered DomalnAPMX-203Y-A214gly - 297-526 Ordered Domain;

N-Disordered Domain - 146-247 Ordered Domain - 297-526 Ordered Domain;

N-Disordered Domain - 146-247 Ordered Domaln-203Y - 297-526 Ordered Domain;

N-Dlsordered Domain - 146-247 Ordered Domaln-A214gly - 297-526 Ordered Domain;

N-Disordered Domain - 146-247 Ordered Domain-203Y-A214gly - 297-526 Ordered Domain;

N-Disordered Domain - 140-247 Ordered Domain - 297-526 Ordered Domain-329Y;

N-Disordered Domain - 140-247 Ordered Domain-203Y - 297-526 Ordered Domaln-329Y;

N-Disordered Domain - 140-247 Ordered Domain-A214gly - 297-526 Ordered Domain-329Y;

N-Disordered Domain - 140-247 Ordered Domain-203Y-A214gly - 297-526 Ordered Domain-329Y;

N-Disordered Domain - 140-247 Ordered DomainAPMX - 297-526 Ordered Domain-329Y;

N-Disordered Domain - 140-247 Ordered DomainAPMX-203Y - 297-526 Ordered Domain-329Y;

N-Disordered Domain - 140-247 Ordered DomalnAPMX-A214gly - 297-526 Ordered Domaln-329Y;

N-Disordered Domain - 140-247 Ordered DomainAPMX-203Y-A214gly - 297-526 Ordered Domain 329Y;

N-Disordered Domain - 146-247 Ordered Domain - 297-526 Ordered Domain-329Y;

N-Disordered Domain - 146-247 Ordered Domain-203Y - 297-526 Ordered Domain-329Y;

N-Disordered Domain - 146-247 Ordered Domain-A214gly - 297-526 Ordered Domain-329Y;

N-Disordered Domain - 146-247 Ordered Domain-203Y-A214gly - 297-526 Ordered Domain-329Y;

N-Disordered Domain - 140-247 Ordered Domain - 297-526 Ordered Domain A297gly;

N-Disordered Domain - 140-247 Ordered Domain-203Y - 297-526 Ordered DomainA297gly;

N-Disordered Domain - 140-247 Ordered Domain-A214gty - 297-526 Ordered Domain A297gly; N-Disordered Domain - 140-247 Ordered Domain-203Y-A214gly - 297-526 Ordered Domain A297gly;

N-Disordered Domain - 140-247 Ordered DomalnAPMX - 297-526 Ordered Domain A297gly;

N-Disordered Domain - 140-247 Ordered DomainAPMX-203Y - 297-526 Ordered Domain A297gly;

N-Disordered Domain - 140-247 Ordered DomainAPMX-A214gly - 297-526 Ordered Domain A297gly;

N-Disordered Domain • 140-247 Ordered DomainAPMX-203Y-A214gly - 297-526 Ordered Domain A297gly;

N-Disordered Domain - 146-247 Ordered Domain - 297-526 Ordered Domain A297gly;

N-Disordered Domain - 146-247 Ordered Domain-203Y - 297-526 Ordered Domain A297gly;

N-Disordered Domain - 146-247 Ordered Domain-A214gly - 297-526 Ordered Domain A297gly;

N-Disordered Domain - 146-247 Ordered Domaln-203Y-A214gly - 297-526 Ordered DomainA297gly;

N-Disordered Domain - 140-247 Ordered Domain - 297-526 Ordered Domain-329YA297gly;

N-Dlsordered Domain - 140-247 Ordered Domaln-203Y - 297-526 Ordered Domain-329YA297gly;

N-Disordered Domain - 140-247 Ordered Domaln-A214gly - 297-526 Ordered Domain-329YA297gly;

N-Disordered Domain - 140-247 Ordered Domaln-203Y-A214gly - 297-526 Ordered Domain 329YA297gly;

N-Disordered Domain - 140-247 Ordered DomalnAPMX - 297-526 Ordered Domain-329YA297gly;

N-Dlsordered Domain - 140-247 Ordered DomalnAPMX-203Y - 297-526 Ordered Domain-329YA297gly;

N-Disordered Domain - 140-247 Ordered DomainAPMX-A214gly - 297-526 Ordered Domain-329YA297gly;

N-Disordered Domain - 140-247 Ordered DomalnAPMX-203Y-A214gly - 297-526 Ordered Domaln-329YA297gly;

N-Disordered Domain - 146-247 Ordered Domain - 297-526 Ordered Domaln-329YA297gly;

N-Dlsordered Domain - 146-247 Ordered Domain-203Y - 297-526 Ordered Domain-329YA297gly;

N-Disordered Domain - 146-247 Ordered Domain-A214gly - 297-526 Ordered Domain-329YA297gly; or

N-Disordered Domain - 146-247 Ordered Domaln-203Y-A214gly - 297-526 Ordered Domain-329YA297gly.

Rh5 ordered domains with N and L Disordered Domains Constructs

N-Dlsordered Domain - 140-247 Ordered Domain - L-Disordered Domain - 297-526 Ordered Domain;

N-Dlsordered Domain - 140-247 Ordered Domain-203Y - L-Disordered Domain - 297-526 Ordered Domain;

N-Disordered Domain - 140-247 Ordered Domain-A214gly - L-Disordered Domain - 297-526 Ordered Domain;

N-Disordered Domain - 140-247 Ordered Domaln-203Y-A214gly - L-Disordered Domain - 297-526 Ordered Domain;

N-Dlsordered Domain - 140-247 Ordered DomalnAPMX - L-Disordered Domain - 297-526 Ordered Domain;

N-Disordered Domain - 140-247 Ordered DomainAPMX-203Y - L-Disordered Domain - 297-526 Ordered Domain;

N-Disordered Domain - 140-247 Ordered DomainAPMX-A214gly - L-Disordered Domain - 297-526 Ordered Domain;

N-Disordered Domain - 140-247 Ordered DomainAPMX-203Y-A214gly - L-Disordered Domain - 297-526 Ordered Domain;

N-Disordered Domain - 146-247 Ordered Domain - L-Disordered Domain - 297-526 Ordered Domain;

N-Disordered Domain - 146-247 Ordered Domain-203Y - L-Disordered Domain - 297-526 Ordered Domain;

N-Disordered Domain - 146-247 Ordered Domain-A214gly - L-Disordered Domain - 297-526 Ordered Domain;

N-Disordered Domain - 146-247 Ordered Domain-203Y-A214gly - L-Disordered Domain - 297-526 Ordered Domain;

N-Disordered Domain - 140-247 Ordered Domain - L-Disordered Domain - 297-526 Ordered Domain-329Y;

N-Disordered Domain - 140-247 Ordered Domain-203Y - L-Disordered Domain - 297-526 Ordered Domain-329Y;

N-Disordered Domain - 140-247 Ordered Domain-A214gly - L-Disordered Domain - 297-526 Ordered Domain-329Y; N-Disordered Domain - 140-247 Ordered Do ma in-203 Y-A214gly - L-Disordered Domain - 297-526 Ordered Domain- 329Y; N-Disordered Domain - 140-247 Ordered DomainAPMX - L-Disordered Domain - 297-526 Ordered Domain-329Y; N-Disordered Domain - 140-247 Ordered DomainAPMX-203Y - L-Disordered Domain - 297-526 Ordered Domain-

329Y;

N-Disordered Domain - 140-247 Ordered DomainAPMX-A214gly - L-Disordered Domain - 297-526 Ordered Domain- 329Y;

N-Disordered Domain - 140-247 Ordered DomainAPMX-203Y-A214gly - L-Disordered Domain - 297-526 Ordered Domain-329Y;

N-Disordered Domain - 146-247 Ordered Domain - L-Disordered Domain - 297-526 Ordered Domain-329Y; N-Disordered Domain - 146-247 Ordered Domaln-203Y - L-Disordered Domain - 297-526 Ordered Domain-329Y; N-Disordered Domain - 146-247 Ordered Domaln-A214gly - L-Disordered Domain - 297-526 Ordered Domaln-329Y; N-Disordered Domain - 146-247 Ordered Domain-203 Y-A214gly - L-Disordered Domain - 297-526 Ordered Domain- 329Y;

N-Disordered Domain - 140-247 Ordered Domain - L-Disordered Domain - 297-526 Ordered Domain A297gly; N-Disordered Domain - 140-247 Ordered Domain-203Y - L-Disordered Domain - 297-526 Ordered DomainA297gly; N-Disordered Domain - 140-247 Ordered Domain-A214gly - L-Disordered Domain - 297-526 Ordered Domain A297gly;

N-Disordered Domain - 140-247 Ordered Domaln-203Y-A214gly - L-Disordered Domain - 297-526 Ordered Domain A297gly;

N-Disordered Domain - 140-247 Ordered DomainAPMX - L-Disordered Domain - 297-526 Ordered Domain A297gly; N-Disordered Domain - 140-247 Ordered DomalnAPMX-203Y - L-Disordered Domain - 297-526 Ordered Domain

A297gly;

N-Disordered Domain - 140-247 Ordered DomalnAPMX-A214gly - L-Disordered Domain - 297-526 Ordered Domain A297gly;

N-Dlsordered Domain - 140-247 Ordered DomalnAPMX-203Y-A214gly - L-Disordered Domain - 297-526 Ordered Domain A297gly;

N-Disordered Domain - 146-247 Ordered Domain - L-Disordered Domain - 297-526 Ordered Domain A297gly;

N-Disordered Domain - 146-247 Ordered Domain-203Y - L-Disordered Domain - 297-526 Ordered Domain A297gly;

N-Disordered Domain - 146-247 Ordered Domain-A214gly - L-Disordered Domain - 297-526 Ordered Domain A297gly;

N-Disordered Domain - 146-247 Ordered Domain-203Y-A214gly - L-Disordered Domain - 297-526 Ordered DomainA297gly;

N-Disordered Domain - 140-247 Ordered Domain - L-Disordered Domain - 297-526 Ordered Domain-329YA297gly;

N-Disordered Domain - 140-247 Ordered Domain-203Y - L-Disordered Domain - 297-526 Ordered Domain-

329YA297gly;

N-Disordered Domain - 140-247 Ordered Domain-A214gly - L-Disordered Domain - 297-526 Ordered Domain- 329YA297gly;

N-Disordered Domain - 140-247 Ordered Domain-203Y-A214gly - L-Disordered Domain - 297-526 Ordered Domain- 329YA297gly;

N-Disordered Domain - 140-247 Ordered DomainAPMX - L-Disordered Domain - 297-526 Ordered Domain-

329YA297gly; N-Disordered Domain - 140-247 Ordered DomainAPMX-203Y - L-Disordered Domain - 297-526 Ordered Domain-

329YA297gly;

N-Disordered Domain - 140-247 Ordered DomainAPMX-A214gly - L-Disordered Domain - 297-526 Ordered Domain- 329YA297gly;

N-Disordered Domain • 140-247 Ordered DomainAPMX-203Y-A214gly - L-Disordered Domain - 297-526 Ordered Domain-329YA297gly;

N-Disordered Domain - 146-247 Ordered Domain - L-Disordered Domain - 297-526 Ordered Domain-329YA297gly;

N-Disordered Domain - 146-247 Ordered Domain-203Y - L-Disordered Domain - 297-526 Ordered Domain-

329YA297gly;

N-Disordered Domain - 146-247 Ordered Domaln-A214gly - L-Disordered Domain - 297-526 Ordered Domaln- 329YA297gly; or

N-Disordered Domain - 146-247 Ordered Domaln-203Y-Δ214gly - L-Disordered Domain - 297-526 Ordered Domain- 329YA297gly.

[0534] In some embodiments, a Plasmodium polypeptide construct described herein includes one or more antigenic portions of a Rh5 polypeptide as described above and also includes a secretory signal (sec). In some embodiments, a secretory signal is a viral secretory signal ( Viralsec) In some embodiments, a secretory signal Is a viral secretory signal Is a HSV-1gD secretory signal (HSV-1gDsec').

[0535] In some embodiments, a Plasmodium polypeptide construct described herein Includes one or more antigenic portions of a Rh5 polypeptide as described above and also includes a transmembrane domain (TMD). In some embodiments, a transmembrane domain Is an HSV-1 transmembrane domain (HSV -1TMD).

[0536] In some embodiments, a Plasmodium polypeptide construct described herein Includes one or more antigenic portions of a Rh5 polypeptide as described above and also includes a multimerization domain (multimerization).

[0537] In some embodiments, a Plasmodium polypeptide construct described herein includes one or more antigenic portions of a Rh5 polypeptide as described above and also includes a self-assembling nanoparticle domain (self-assem).

[0538] In some embodiments, a Plasmodium polypeptide construct described herein further Includes a linker. A linker may be included between any two domains of a Plasmodium polypeptide construct In some embodiments, a linker is included after a Rh5 domain of a Rh5 polypeptide construct

[0539] In some embodiments, a Rh5 polypeptide construct can have the following structure:

Rh5 ordered domain Construct; sec - Rh5 ordered domain Construct;

Viralsec - Rh5 ordered domain Construct;

HSV-lgDsec. - Rh5 ordered domain Construct; Rh5 ordered domain Construct - TMD; sec - Rh5 ordered domain Construct - TMD;

Viralsec. - Rh5 ordered domain Construct - TMD;

HSV-lgDsec - Rh5 ordered domain Construct - TMD; Rh5 ordered domain Construct - HSV -1TMD; sec - Rh5 ordered domain Construct - HSV -1TMD; Viralsec - Rh5 ordered domain Construct - HSV -1TMD,- HSV-1gDsec - Rh5 ordered domain Construct - HSV -1TMD; Rh5 ordered domain Construct - multimerization; sec - Rh5 ordered domain Construct - multimerization;

Viralsec - Rh5 ordered domain Construct - multimerization;

HSV-1gDsec. - Rh5 ordered domain Construct - multimerization; Rh5 ordered domain Construct - self-assem; sec - Rh5 ordered domain Construct - self-assem;

Viralsec - Rh5 ordered domain Construct - self-assem;

HSV-1gDsec - Rh5 ordered domain Construct - self-assem; Rh5 ordered domain Construct - Linker - TMD; sec - Rh5 ordered domain Construct - Linker - TMD;

Viralsec - Rh5 ordered domain Construct - Linker - TMD;

HSV-1gDsec - Rh5 ordered domain Construct - Linker - TMD; Rh5 ordered domain Construct - Linker - HSV -1TMD; sec - Rh5 ordered domain Construct - Linker - HSV -1TMD;

Viralsec - Rh5 ordered domain Construct - Linker - HSV -1TMD; HSV-1gDsec - Rh5 ordered domain Construct - Linker - HSV -1TMD; Rh5 ordered domain Construct - Linker - multimerization; sec - Rh5 ordered domain Construct - Linker - multimerization;

Viralsec - Rh5 ordered domain Construct - Linker - multimerization; HSV-1gDsec - Rh5 ordered domain Construct - Linker - multimerization; Rh5 ordered domain Construct - Linker - self-assem; sec - Rh5 ordered domain Construct - Linker - self-assem;

Viralsec - Rh5 ordered domain Construct - Linker - self-assem;

HSV-1gDsec - Rh5 ordered domain Construct - Linker - self-assem; Rh5 ordered domain with Linker Construct; sec - Rh5 ordered domain with Linker Construct;

Viralsec - Rh5 ordered domain with Linker Construct;

HSV-1gDsec - Rh5 ordered domain with Linker Construct; Rh5 ordered domain with Linker Construct - TMD; sec - Rh5 ordered domain with Linker Construct - TMD;

Viralsec - Rh5 ordered domain with Linker Construct - TMD;

HSV-1gDsec - Rh5 ordered domain with Linker Construct - TMD; Rh5 ordered domain with Linker Construct - HSV -1TMD; sec - Rh5 ordered domain with Linker Construct - HSV -1TMD;

Viralsec - Rh5 ordered domain with Linker Construct - HSV -1TMD; HSV-1gDsec - Rh5 ordered domain with Linker Construct - HSV -1TMD; Rh5 ordered domain with Linker Construct - multimerization; sec - Rh5 ordered domain with Linker Construct - multimerization; Viralsec - Rh5 ordered domains with L-Disordered Domain Construct - self-assem;

HSV-1gDsec - Rh5 ordered domains with L-Disordered Domain Construct - self-assem; Rh5 ordered domains with L-Disordered Domain Construct - Linker - TMD; sec - Rh5 ordered domains with L-Disordered Domain Construct - Linker - TMD;

Viralsec - Rh5 ordered domains with L-Disordered Domain Construct - Linker - TMD;

HSV-1gDsec - Rh5 ordered domains with L-Disordered Domain Construct - Linker - TMD; Rh5 ordered domains with L-Disordered Domain Construct - Linker - HSV-1TMD; sec - Rh5 ordered domains with L-Disordered Domain Construct - Linker - HSV -1TMD;

Viralsec - Rh5 ordered domains with L-Disordered Domain Construct - Linker - HSV-1TMD;

HSV-1gDsec. - Rh5 ordered domains with L-Disordered Domain Construct - Linker - HSV-1TMD; Rh5 ordered domains with L-Disordered Domain Construct - Linker - multimerization; sec - Rh5 ordered domains with L-Disordered Domain Construct - Linker - multimerization;

Viralsec - Rh5 ordered domains with L-Disordered Domain Construct - Linker - multimerization;

HSV-1gDsec - Rh5 ordered domains with L-Disordered Domain Construct - Linker - multimerization; Rh5 ordered domains with L-Disordered Domain Construct - Linker - self-assem; sec - Rh5 ordered domains with L-Disordered Domain Construct - Linker - self-assem;

Viralsec - Rh5 ordered domains with L-Disordered Domain Construct - Linker - self-assem;

HSV-1gDsec - Rh5 ordered domains with L-Disordered Domain Construct - Linker - self-assem; Rh5 ordered domains with N-Disordered Domain Construct; sec - Rh5 ordered domains with N-Disordered Domain Construct;

Viralsec - Rh5 ordered domains with N-Disordered Domain Construct,

HSV-1gDsec - Rh5 ordered domains with N-Disordered Domain Construct, Rh5 ordered domains with N-Disordered Domain Construct - TMD; sec - Rh5 ordered domains with N-Disordered Domain Construct - TMD;

Viralsec - Rh5 ordered domains with N-Disordered Domain Construct - TMD;

HSV-1gDsec - Rh5 ordered domains with N-Disordered Domain Construct - TMD; Rh5 ordered domains with N-Disordered Domain Construct - HSV-1TMD; sec - Rh5 ordered domains with N-Disordered Domain Construct - HSV-1TMD;

Viralsec - Rh5 ordered domains with N-Disordered Domain Construct - HSV-1TMD;

HSV-1gDsec - Rh5 ordered domains with N-Disordered Domain Construct - HSV-1TMD; Rh5 ordered domains with N-Disordered Domain Construct - multimerization; sec - Rh5 ordered domains with N-Disordered Domain Construct - multimerization;

Viralsec - Rh5 ordered domains with N-Disordered Domain Construct - multimerization;

HSV-1gDsec - Rh5 ordered domains with N-Disordered Domain Construct - multimerization; Rh5 ordered domains with N-Disordered Domain Construct - self-assem; sec - Rh5 ordered domains with N-Disordered Domain Construct - self-assem;

Viralsec - Rh5 ordered domains with N-Disordered Domain Construct - self-assem;

HSV-1gDsec - Rh5 ordered domains with N-Disordered Domain Construct - self-assem; Rh5 ordered domains with N-Disordered Domain Construct - Linker - TMD; sec - Rh5 ordered domains with N-Disordered Domain Construct - Linker - TMD; Viralsec - Rh5 ordered domains with N and L Disordered Domains Construct - Linker - HSV -1TMD;

HSV-1gDsec - Rh5 ordered domains with N and L Disordered Domains Construct - Linker - HSV -1TMD; Rh5 ordered domains with N and L Disordered Domains Construct - Linker - multimerization; sec - Rh5 ordered domains with N and L Disordered Domains Construct - Linker - multimerization;

Viralsec - Rh5 ordered domains with N and L Disordered Domains Construct - Linker - multimerization;

HSV-1gDsec - Rh5 ordered domains with N and L Disordered Domains Construct - Linker - multimerization; Rh5 ordered domains with N and L Disordered Domains Construct - Linker • self-assem; sec - Rh5 ordered domains with N and L Disordered Domains Construct - Linker - self-assem;

Viralsec - Rh5 ordered domains with N and L Disordered Domains Construct - Linker - self-assem; or HSV-1gDsec - Rh5 ordered domains with N and L Disordered Domains Construct - Linker - self-assem.

[0540] Table 7 provides exemplary Rh5 constructs. The different domains in the Rh5 constructs in Table 7 are further labeled as follows: secretion signal sequence is underlined, anker sequence is in italics, and additional domains are in bold (transmembrane, multimerization, self-assembling domain)

Table 7: Amino Acid Sequences Encoded by Exemplary Rh5 RNA Constructs as Described Herein

[0541] In some embodiments, a Rh5 construct described herein has an amino acid sequence provided in Table 7. In some embodiments, a Rh5 construct comprises an amino add sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino add sequence of Table 7.

[0542] In some embodiments, a Rh5 construct conyrises or consists of an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino acid sequence of SEQ ID NO: 98. In some embodiments, a Rh5 construct comprises or consists of an amino acid sequence of SEQ ID NO: 98.

[0543] In some embodiments, a Rh5 construct comprises or consists of an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino add sequence of SEQ ID NO: 99. In some embodiments, a Rh5 construct comprises or consists of an amino acid sequence of SEQ ID NO: 99.

[0544] In some embodiments, a Rh5 construct comprises or consists of an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino add sequence of SEQ ID NO: 100. In some embodiments, a Rh5 construct comprises or consists of an amino add sequence of SEQ ID NO: 100.

[0545] In some embodiments, a Rh5 construct comprises or consists of an amino add sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino add sequence of SEQ ID NO: 101. In some embodiments, a Rh5 construct comprises or consists of an amino add sequence of SEQ ID NO: 101.

[0546] In some embodiments, a Rh5 construct comprises or consists of an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino add sequence of SEQ ID NO: 102. In some embodiments, a Rh5 construct comprises or consists of an amino acid sequence of SEQ ID NO: 102.

[0547] In some embodiments, a Rh5 construct comprises or consists of an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino add sequence of SEQ ID NO: 103. In some embodiments, a Rh5 construct comprises or consists of an amino add sequence of SEQ ID NO: 103.

[0548] In some embodiments, a Rh5 construct comprises or consists of an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino add sequence of SEQ ID NO: 104. In some embodiments, a Rh5 construct comprises or consists of an amino add sequence of SEQ ID NO: 104.

[0548] In some embodiments, a Rh5 construct comprises or consists of an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino add sequence of SEQ ID NO: 105. In some embodiments, a Rh5 construct comprises or consists of an amino add sequence of SEQ ID NO: 105.

[0550] In some embodiments, a Rh5 construct comprises or consists of an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino add sequence of SEQ ID NO: 106. In some embodiments, a Rh5 construct comprises or consists of an amino add sequence of SEQ ID NO: 106. [0551] In some embodiments, a Rh5 construct comprises or consists of an amino add sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino add sequence of SEQ ID NO: 107. In some embodiments, a Rh5 construct comprises or consists of an amino add sequence of SEQ ID NO: 107.

[0552] In some embodiments, a Rh5 construct comprises or consists of an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino add sequence of SEQ ID NO: 108. In some embodiments, a Rh5 construct comprises or consists of an amino add sequence of SEQ ID NO: 108.

[0553] In some embodiments, a Rh5 construct comprises or consists of an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino add sequence of SEQ ID NO: 109. In some embodiments, a Rh5 construct comprises or consists of an amino add sequence of SEQ ID NO: 109.

Table 8: DNA Sequences Corresponding to Exemplary Rh5 RNA Constructs as Described Herein Table 9: RNA Sequences Corresponding to Exemplary Rh5 RNA Constructs as Described Herein

SEQ RN* Construct Sequence (RNA) ID NO:

225 1 AUGGGCGGAGCUGCUGCUAGACUGGGAGCCGUGAUCCUGUUCGUGGUUAUCGUG

GGACUGCAUGGCGUGCGGGGAGAGAACGCCAUCAAAAAGACCAAGAAUCAAGAGAA

CAACCUGGCUCUGCUGCCCAUCAAGAGCACCGAGGAAGAAAAGGACGACAUCAAGA

ACGGCAAGGAUAUCAAGAAAGAGAUCGACAACGACAAAGAGAACAUCAAGACCAACA

ACGCCAAGGACCACAGCACCUACAUCAAGUCCUACCUGAACACCAACGUGAACGACG

GCCUGAAGUACCUGUUCAUCCCCAGCCACAACAGCUUUAUCAAAAAGUACAGCGUG

ULICAACCAGAUCAACGACGGCAUGCUGCUGAACGAGAAGAACGACGUGAAGAACAA

CGAGGACUACAAGAACGUCGACUAUAAGAAUGUGAACUUCCUCCAGUACCACUUCA

AAGAGCUGAGCAACUACAAUAUCGCCAACAGCAUCGACAUCCUGCAAGAGAAAGAG

GGCCACCUGGACUUCGUGAUCAUCCCUCACUACACCUUUCUGGACUACUACAAGCA

CCUGAGCUACAACUCCAUCUACCACAAGUCCUCCACCUACGGCAAGUGUAUCGCCG

UGGACGCCUUCAUCAAGAAGAUUAACGAGGCCUACGACAAAGUGAAGUCCAAGUGC

AACGACAUUAAGAAUGACCUGAUCGCCACGAUCAAGAAGCUGGAACACCCCUACGA

UAUCAACAACAAGAACGAUGACAGCUACCGCUACGACAUCAGCGAGGAAAUCGACG

ACAAGUCCGAGGAAACAGACGACGAGACAGAGGAAGUGGAAGAUAGCAUCCAGGAC

ACCGACAGCAAUCACGCCCCUAGCAACAAGAAAAAGAACGACCUCAUGAACCGGGCC

UUCAAGAAAAUGAUGGACGAGUACAACACCAAGAAGAAAAAGCUGAUCAAGUGCAU

CAAAAACCACGAGAACGACUUCAACAAGAUCUGCAUGGACAUGAAGAACUACGGCA

CCAACCUGUUCGAGCAGCUGUCCUGCUACAACAACAACUUCUGCAACACGAACGGC

AUCCGCUACCACUACGAUGAGUACAUCCACAAGCUGAUCCUGAGCGUCAAGAGCAA

GAACCUGAACAAGGACCUGUCCGACAUGACCAACAUCCUCCAGCAGAGCGAACUGC

UGCUGACCAAUCUGAACAAGAAGAUGGGCUCCUACAUCUACAUCGACACCAUCAAG

UUCAUCCACAAAGAGAUGAAGCACAUCUUCAACCGGAUCGAGUACCACACGAAGAU

CAUCAACGACAAGACUAAGAUCAUCCAGGAUAAGAUCAAGCUGAACAUCUGGCGGA

CCUUCCAGAAGGACGAGCUGCUGAAGCGCAUCCUGGACAUGAGCAACGAGUACAGC

CUGUUUAUCACCAGCGACCACCUGAGACAGAUGCUCUACAACACCUUCUACAGCAA

AGAGAAGCACCUCAACAACAUCUUCCACCACCUGAUCUACGUGCUCCAGAUGAAGU

UCAACGACGUGCCGAUCAAGAUGGAAUACUUCCAGACCUACAAGAAGAACAAACCC

CUGACACAGUGAUAA _

227 2 AUGGGCGGAGCUGCUGCUAGACUGGGAGCCGUGAUCCUGUUCGUGGUUAUCGUG

GGACUGCAUGGCGUGCGGGGCAAGAACGUGAACUUCCUCCAGUACCACUUCAAAGA

GCUGAGCAACUACAAUAUCGCCAACAGCAUCGACAUCCUGCAAGAGAAAGAGGGCC

ACCUGGACUUCGUGAUCAUCCCUCACUACACCUUUCUGGACUACUACAAGCACCUG

AGCUACAACUCCAUCUACCACAAGAGCAGCACCUACGGCAAGUAUAUCGCCGUGGA

CGCCUUCAUCAAGAAGAUCAACGAGACAUACGACAAAGUGAAGUCCAAGUGCAACG

ACAUCAAGAACGACCUGAUCGCCACGAUCAAGAAGCUGGAACACCCCUACGAUAUC

AACAACAAGAACCGGACCUUCAAGAAAAUGAUGGACGAGUACAACACCAAGAAGAAA

AAGCUGAUCAAGUGCAUCAAGAAUCACGAGAACGACUUCAACAAGAUCUGCAUGGA

CAUGAAGAACUACGGCACCAACCUGUUCGAGCAGCUQJCCUGCUACAACAACAACU

UCUGCAACACCAACGGCAUCCGCUACCACUACGAUGAGUACAUCCACAAGCUGAUC

CUGAGCGUCAAGUCCAAGAACCUGAACAAGGACCUGUCCGACAUGACCAACAUCCU

CCAGCAGAGCGAACUGCUGCUGACCAAUCUGAACAAGAAGAUGGGCAGCUACAUCU

ACAUCGACACCAUCAAGUUCAUCCACAAAGAGAUGAAGCACAUCUUCAACCGGAUC

GAGUACCACACGAAGAUCAUCAACGACAAGACAAAGAUCAUCCAGGAUAAGAUCAA

GCUGAACAUCUGGCGCACCUUCCAGAAGGACGAGCUGCUGAAGCGCAUCCUGGACA

UGAGCAACGAGUACAGCCUGUUCAUCACCAGCGACCACCUGAGACAGAUGCUGUAC

AAUACCUUCUACAGCAAAGAGAAGCACCUCAACAACAUCUUCCACCACCUGAUCUAC

GUGCUCCAGAUGAAGUUCAACGACGUGCCCAUCAAGAUGGAAUACUUCCAGACCUA

CAAGAAGAACAAACCCCUGACACAGGGCAGCCCUGGCUCUGGUAGUGGCUCUGGCU

AUAUCCCCGAGGCUCCUAGAGAUGGCCAGGCCUAUGUUCGGAAGGAUGGCGAAUG

GGUGCUGCUGAGCACAUUCCUGUGAUAA _

229 6 AUGGGCGGAGCUGCUGaJAGACUGGGAGCCGUGAUCCUGUUCGUGGUUAUCGUG

GGACUGCAUGGCGUGCGGGGCAAGAACGUGAACUUCCUCCAGUACCACUUCAAAGA GCUGAGCAACUACAAUAUCGCCAACAGCAUCGACAUCCUGCAAGAGAAAGAGGGCC

[0555] In some embodiments, a Rh5 construct described herein has a nucleic acid sequence provided in Table

9. In some embodiments, a Rh5 construct comprises a nudek: add sequence having at least 99%, 98%, 97%, 96%,

95%, 90%, 85%, or 80% identity to a nucleic add sequence of Table 9.

2. CyRPA constructs

[0556] In some embodiments, the Plasmodium polypeptide construct described herein includes a CyRPA polypeptide or antigenic portion thereof from Plasmodium falciparum, preferably from Plasmodium falciparum isolate

3D7. In some embodiments, the Plasmodium polypeptide construct described herein indudes an antigenic portion of CyRPA from Plasmodium falciparum, preferably from Plasmodium falciparum isolate 3D7 (referred to herein as a "CyRPA construct" or “CyRPA domain").

[0557] In some embodiments, an antigenic portion of a Plasmodium CyRPA polypeptide in a CyRPA construct comprises the amino add sequence of positions 30-362 of SEQ ID NO: 3, or an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the amino add sequence of positions 30-362 of SEQ ID NO: 3. In some embodiments, an antigenic portion of a Plasmodium CyRPA polypeptide in a CyRPA construct comprises the amino add sequence of positions 30-362 of SEQ ID NO: 3, or the amino add sequence of positions 30- 362 of SEQ ID NO: 3 having 1, 2, 3, 4, or 5 amino acid substitutions. In some embodiments, an antigenic portion of a Plasmodium CyRPA polypeptide in a CyRPA construct comprises or consists of a sequence according to SEQ ID NO: 133 or a variant thereof (e.g., a glycosylation variant). In some embodiments, an antigenic portion of a Plasmodium CyRPA polypeptide in a CyRPA construct comprises or consists of a sequence with at least 85% sequence identity to SEQ ID NO: 133 or a variant thereof (e.g., a glycosylation variant).

[0558] In some embodiments, an antigenic portion of a Plasmodium CyRPA polypeptide In a CyRPA construct comprises or consists of an amino add sequence of SEQ ID NO: 133 that has been modified to include one or more amino acid substitutions that prevent glycosylation at amino add positions 145, 332, and/or 338 (as numbered according to SEQ ID NO: 3). In some embodiments, an antigenic portion of a Plasmodium CyRPA polypeptide in a CyRPA construct comprises or consists of an amino acid sequence of SEQ ID NO: 133 that has two amino add substitutions that prevent glycosylation at two positions selected from: 145, 332, and 338 (as numbered according to SEQ ID NO: 3). In some embodiments, an antigenic portion of a Plasmodium CyRPA polypeptide In a CyRPA construct comprises or consists of an amino add sequence of SEQ ID NO: 134.

[0559] In some embodiments, an antigenic portion of a Plasmodium CyRPA polypeptide in a CyRPA construct comprises one, two, or three N-llnked glycosylation sites. In some embodiments, a Plasmodium CyRPA antigenic portion comprises an amino add substitution at one or more N-linked glycosylation sites, where the amino acid substitution prevents glycosylation. In some embodiments, a Plasmodium CyRPA antigenic portion in a CyRPA construct comprises two amino add substitutions at two N-llnked glycosylation sites, where the amino add substitution prevents glycosylation. In some embodiments, a Plasmodium CyRPA antigenic portion In a CyRPA construct comprises an amino acid substitutions at all N-linked glycosylation sites, where the amino add substitution prevents glycosylation.

[0560] In some embodiments an antigenic portion of a Plasmodium CyRPA polypeptide in a CyRPA construct comprises or consists of an amino acid sequence that is at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 133 and comprises one or more substitutions selected from: N145Q, N332Q, and N338Q.

[0561] In some embodiments, a Plasmodium CyRPA antigenic portion in a CyRPA construct comprises an asparagine at position 116, position 293, position 309, or a combination thereof, as numbered according to SEQ ID NO: 133. In some embodiments, a Plasmodium CyRPA antigenic portion in a CyRPA construct comprises a glutamine at position 116, position 293, position 309, or a combination thereof, as numbered according to SEQ ID NO: 133. In some embodiments, a Plasmodium CyRPA antigenic portion in a CyRPA construct comprises a glutamine at position 116, position 293, and position 309, as numbered according to SEQ ID NO: 133. In some embodiments, a Plasmodium CyRPA antigenic portion in a CyRPA construct comprises or consists of an amino acid sequence of SEQ ID NO: 134. In some embodiments, a Plasmodium CyRPA antigenic portion in a CyRPA construct comprises or consists of an amino add sequence of SEQ ID NO: 170. [0562] In some embodiments, a Plasmodium polypeptide construct described herein further includes one or more linkers. In some embodiments, a Plasmodium polypeptide construct described herein further includes one or more linkers between one or more regions and/or domains. In some embodiments, a linker comprises or consists of an amino acid sequence according to SEQ ID NO: 86. In some embodiments, a linker comprises or consists of an amino add sequence according to SEQ ID NO: 138.

[0563] In some embodiments, a Plasmodium polypeptide construct described herein further indudes one or more secretory signals. In some embodiments, a secretory signal is a Plasmodium CyRPA secrtary signal (CyRPAsec). In some embodiments, a secretory signal comprises or consists of an amino acid sequence according to SEQ ID NO: 187. In some embodiments, a secretory signal is a viral secretory signal. In some embodiments, a secretory signal Is an HSV glycoprotein D (gD) secretory signal. In some embodiments, an HSV glycoprotein D (gD) secretory signal comprises or consists of an amino acid sequence according to SEQ ID NO: 42.

[0564] In some embodiments, a Plasmodium polypeptide construct described herein further comprises one or more transmembrane regions. In some embodiments, a transmembrane region is a heterologous transmembrane region. In some embodiments, a transmembrane region comprises or consists of a viral transmembrane region. In some embodiments, a transmembrane region comprises or consists of an HSV transmembrane region. In some embodiments, a transmembrane region comprises or consists of an HSV gD transmembrane region. In some embodiments, a transmembrane region comprises or consists of an amino add sequence according to SEQ ID NO: 75.

[0565] In some embodiments, a Plasmodium polypeptide construct described herein further comprises one or more tags. In some embocflments, a tag Is a HIBit tag. In some embodiments, a tag comprises or consists of a sequence according to SEQ ID NO: 139.

[0566] In some embodiments, a Plasmodium polypeptide construct can have the following structure:

CyRPA construct sec - CyRPA construct;

Viralsec - CyRPA construct;

HSV-1gDsec. - CyRPA construct;

CyRPAsec - CyRPA construct;

CyRPA construct - TMD; sec - CyRPA construct - TMD;

Viralsec - CyRPA construct - TMD;

HSV-1gDsec. - CyRPA construct - TMD;

CyRPAsec - CyRPA construct - TMD;

CyRPA construct - HSV -1TMD; sec - CyRPA construct - HSV -1TMD;

Viralsec - CyRPA construct - HSV -1TMD;

HSV-1gDsec. - CyRPA construct - HSV -1TMD;

CyRPAsec - CyRPA construct - HSV -1TMD;

CyRPA construct - multimerization; sec - CyRPA construct - multimerization;

Viralsec - CyRPA construct - multimerization;

[0567] In some embodiments, a CyRPA construct described herein has an amino acid sequence provided in Table 10. In some embodiments, a CyRPA construct comprises an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% Identity to an amino add sequence of Table 10.

[0568] In some embodiments, a CyRPA construct comprises or consists of an amino add sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% Identity to an amino add sequence of SEQ ID NO: 140. In some embodiments, a CyRPA construct comprises or consists of an amino acid sequence of SEQ ID NO: 140.

[0569] In some embodiments, a CyRPA construct comprises or consists of an amino add sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino add sequence of SEQ ID NO: 141. In some embodiments, a CyRPA construct comprises or consists of an amino acid sequence of SEQ ID NO: 141.

[0570] In some embodiments, a CyRPA construct comprises or consists of an amino add sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% Identity to an amino add sequence of SEQ ID NO: 142. In some embodiments, a CyRPA construct comprises or consists of an amino add sequence of SEQ ID NO: 142.

[0571] In some embodiments, a CyRPA construct comprises or consists of an amino add sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% Identity to an amino add sequence of SEQ ID NO: 143. In some embodiments, a CyRPA construct comprises or consists of an amino acid sequence of SEQ ID NO: 143. [0572] In some embodiments, a CyRPA construct comprises or consists of an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino add sequence of SEQ ID NO: 144. In some embodiments, a CyRPA construct comprises or consists of an amino add sequence of SEQ ID NO: 144.

[0573] In some embodiments, a CyRPA construct comprises or consists of an amino add sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino add sequence of SEQ ID NO: 198. In some embodiments, a CyRPA construct comprises or consists of an amino acid sequence of SEQ ID NO: 198.

[0574] In some embodiments, a CyRPA construct comprises or consists of an amino add sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino add sequence of SEQ ID NO: 199. In some embodiments, a CyRPA construct comprises or consists of an amino acid sequence of SEQ ID NO: 199.

Table 11: DNA Sequences Corresponding to Exemplary CyRPA RNA Constructs as Described Herein TAGGGACTTTCTCAAGGATAACAAAGTGCTGCAAGACGTATCCACCCTGAACGACGA

GTATATCGTGAGCTACGGGAATGATAATAACTTCGCCGAGTGCTACATnTTTTCAAT

AACGAGAACTCCATCCTGATCAAGCCTGAAAAGTACGGCCAGACTACCGCCGGTTGC

TACGCHGGGACCTTCGrrCAAAATTGACGAGCAGCGTACATTGTTCATCTATAGTTCT

AGTCAGGGCATCTACAACATTCACACCATCTATTACGCTAACTACGAGGGAGGCTCT

GGCGGTGGACTTATTGCCGGCGCTGTTGGAGGATCTCTGCTGGCCGCTCTGGrTCAT CTGTGGCATCGTGTACTGGATGCGGCGGCACACACAGAAGGCCCCTAAGAGAATCAG ACTGCCCCACATCAGATGATGA

[0575] In some embodiments, a CyRPA construct described herein has a nucleic add sequence provided In

Table 11. In some embodiments, a CyRPA construct comprises a nucleic acid sequence having at least 99%, 98%,

97%, 96%, 95%, 90%, 85%, or 80% Identity to a nudek add sequence of Table 11.

Table 12: RNA Sequences Corresponding to Exemplary CyRPA RNA Constructs as Described Herein

SEQ RNA Construct Sequence (RNA)

ID

NO:

243 22 AUGAUUAUCCCAUUCCACAAAAAGUUCAUCUCCUUUUUCCAGAUCGUACUGGUGGU ACUGCUUCUGUGCAGAUCCAUUAACUGUGAUUCCCGUCACGUUUUCAUUCGUACC GAGCUGAGCUUUAUCAAGAACAAUGUCCCCUGUAUCCGCGACAUGUUUUUCAUUU ACAAGCGUGAACUGUACAACAUUUGCCUCGAUGACCUUAAGGGCGAGGAGGACGAA ACCCACAUUUACGUGCAAAAGAAAGUGAAAGAUAGCUGGAUCACGCUGAACGACCU GUUCAAGGAGACAGAUCUGACCGGGCGCCCUCAUAUCUUCGCUUACGUGGAUGUG GAGGAGAUUAUCAUUCUCCUGUGUGAAGAUGAGGAGUUCUCUAACCGCAAGAAAG ACAUGACUUGCCACCGGUUCUACUCCAACGACGGGAAGGAGUAUAAUAACUCUGAA AUCACCAUUAGCGACUAUAUCCUCAAGGACAAGCUGCUCAGCUCCUAUGUGUCCCU GCCCCUGAAGAUUGAAAAUCGCGAAUACUUCCUGAUUUGUGGGGUGUCUCCAUAC AAGUUUAAGGAUGACAACAAAAAGGACGAUAUCCUGUGCAUGGCGUCACACGAUAA GGGAGAAACCUGGGGCACCAAGAUCGUCAUCAAGUACGAUAAUUACAAGCUGGGU GUUCAGUACUUUUUCCUGCGUCCAUACAUCUCCAAGAACGAUCUGUCUUUCCACUU UUAUGUCGGAGAUAACAUUAAUAACGUGAAGAACGUGAAUUUUAUUGAGUGCACG CAUGAAAAGGACCUGGAGUUUGUCUGUUCUAAUAGGGACUUUCUCAAGGAUAACA AAGUGCUGCAAGACGUAUCCACCCUGAACGACGAGUAUAUCGUGAGCUACGGGAAU GAUAAUAACUUCGCCGAGUGCUACAUUUUUUUCAAUAACGAGAACUCCAUCCUGAU CAAGCCUGAAAAGUACGGCAACACUACCGCCGGUUGCUACGGUGGGACCUUCGUCA AAAUUGACGAGAACCGUACAUUGUUCAUCUAUAGUUCUAGUCAGGGCAUCUACAAC AUUCACACCAUCUAUUACGCUAACUACGAGUGAUGA

245 23 AUGAUUAUCCCAUUCCACAAAAAGUUCAUCUCCUUUUUCCAGAUCGUACUGGUGGU ACUGCUUCUGUGCAGAUCCAUUAACUGUGAUUCCCGUCACGUUUUCAUUCGUACC GAGCUGAGCUUUAUCAAGAACAAUGUCCCCUGUAUCCGCGACAUGUUUUUCAUUU ACAAGCGUGAACUGUACAACAUUUGCCUCGAUGACCUUAAGGGCGAGGAGGACGAA ACCCACAUUUACGUGCAAAAGAAAGUGAAAGAUAGCUGGAUCACGCUGAACGACCU GUUCAAGGAGACAGAUCUGACCGGGCGCCCUCAUAUCUUCGCUUACGUGGAUGUG GAGGAGAUUAUCAUUCUCCUGUGUGAAGAUGAGGAGUUCUCUAACCGCAAGAAAG ACAUGACUUGCCACCGGUUCUACUCCAACGACGGGAAGGAGUAUCAGAACUCUGAA AUCACCAUUAGCGACUAUAUCCUCAAGGACAAGCUGCUCAGCUCCUAUGUGUCCCU GCCCCUGAAGAUUGAAAAUCGCGAAUACUUCCUGAUUUGUGGGGUGUCUCCAUAC AAGUUUAAGGAUGACAACAAAAAGGACGAUAUCCUGUGCAUGGCGUCACACGAUAA GGGAGAAACCUGGGGCACCAAGAUCGUCAUCAAGUACGAUAAUUACAAGCUGGGU GUUCAGUACUUUUUCCUGCGUCCAUACAUCUCCAAGAACGAUCUGUCUUUCCACUU UUAUGUCGGAGAUAACAUUAAUAACGUGAAGAACGUGAAUUUUAUUGAGUGCACG CAUGAAAAGGACCUGGAGUUUGUCUGUUCUAAUAGGGACUUUCUCAAGGAUAACA

AAGUGCUGCAAGACGUAUCCACCCUGAACGACGAGUAUAUCGUGAGCUACGGGAAU GAUAAUAACUUCGCCGAGUGCUACAUUUUUUUCAAUAACGAGAACUCCAUCCUGAU CAAGCCUGAAAAGUACGGCCAGACUACCGCCGGUUGCUACGGUGGGACCUUCGUCA GGAUGACAACAAAAAGGACGAUAUCCUGUGCAUGGCGUCACACGAUAAGGGAGAAA

CCUGGGGCACCAAGAUCGUCAUCAAGUACGAUAAUUACAAGCUGGGUGUUCAGUAC

UUUUUCCUGCGUCCAUACAUCUCCAAGAACGAUCUGUCUUUCCACUUUUAUGUCG

GAGAUAACAUUAAUAACGUGAAGAACGUGAAUUUUAUUGAGUGCACGCAUGAAAAG

GACCUGGAGUUUGUCUGUUCUAAUAGGGACUUUCUCAAGGAUAACAAAGUGCUGC

AAGACGUAUCCACCCUGAACGACGAGUAUAUCGUGAGCUACGGGAAUGAUAAUAAC

UUCGCCGAGUGCUACAUUUUUUUCAAUAACGAGAACUCCAUCCUGAUCAAGCCUGA

AAAGUACGGCCAGACUACCGCCGGUUGCUACGGUGGGACCUUCGUCAAAAUUGACG

AGCAGCGUACAUUGUUCAUCUAUAGUUCUAGUCAGGGCAUCUACAACAUUCACACC

AUCUAUUACGCUAACUACGAGGGAGGOJCUGGCGGUGGACUUAUUGCCGGCGCUG

UUGGAGGAUCUCUGCUGGCCGCUCUGGUCAUCUGUGGCAUCGUGUACUGGAUGCG

GCGGCACACACAGAAGGCCCCUAAGAGAAUCAGACUGCCCCACAUCAGAUGAUGA

[0576] In some embodiments, a CyRPA construct described herein has a nucleic add sequence provided In Table 12. In some embodiments, a CyRPA construct comprises a nucleic acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to a nudek add sequence of Table 12.

3. P113 constructs

[0577] In some embodiments, the Plasmodium polypeptide construct described herein includes a P113 polypeptide or antigenic portion thereof from Plasmodium faidparum, preferably from Plasmodium falciparum Isolate 3D7. In some embodiments, the Plasmodium polypeptide construct described herein Includes an antigenic portion of P113 from Plasmodium faidparum, preferably from Plasmodium falciparum isolate 3D7 (referred to herein as a "P113 construct” or ”P113 domain").

[0578] In some embodiments, an antigenic portion of a Plasmodium P113 polypeptide in a P113 construct comprises the amino add sequence of positions 23-939 of SEQ ID NO: 6, or an amino add sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the amino add sequence of positions 23-939 of SEQ ID NO: 6. In some embodiments, an antigenic portion of a Plasmodium Pl 13 polypeptide in a P113 construct comprises the amino acid sequence of positions 23-939 of SEQ ID NO: 6, or the amino acid sequence of positions 23-939 of SEQ ID NO: 6 having 1, 2, 3, 4, or 5 amino add substitutions.

[0579] In some embodiments, an antigenic portion of a Plasmodium P113 polypeptide In a P113 construct comprises one, two, three, four, five, six, seven, or eight N-linked glycosylation sites. In some embodiments, an antigenic portion of a Plasmodium P113 polypeptide in a P113 construct comprises an amino add substitution at one or more N -linked glycosylation sites, where the amino acid substitution prevents glycosylation. In some embodiments, an antigenic portion of a Plasmodium Pl 13 polypeptide in a P113 construct comprises two, three, four, five, six, or seven amino acid substitutions at N-linked glycosylation sites, where the amino add substitution prevents glycosylation. In some embodiments, an antigenic portion of a Plasmodium P113 polypeptide in a P113 construct comprises an amino acid substitutions at all N-linked glycosylation sites, where the amino add substitution prevents glycosylation.

[0580] In some embodiments, an antigenic portion of a Plasmodium P113 polypeptide in a Pl 13 construct comprises or consists of an amino add sequence of SEQ ID NO: 135 that has been modified to indude one or more amino add substitutions that prevent glycosylation at amino add positions 207, 268, 317, 360, 661, 697, 779, 876, or 938, as numbered according to SEQ ID NO: 6. In some embodiments, an antigenic portion of a Plasmodium P113 polypeptide in a P113 construct comprises or consists of an amino add sequence of SEQ ID NO: 135 that has been modified to indude two or more amino add substitutions that prevent glycosylation at amino add position selected from 207, 268, 317, 360, 661, 697, 779, 876, and 938, as numbered according to SEQ ID NO: 6. In some embodiments, an antigenic portion of a Plasmodium P113 polypeptide in a P113 construct comprises or consists of an amino acid sequence of SEQ ID NO: 135 that has been modified to include three, four, five, six, seven or more amino add substitutions that prevent glycosylation at amino add position selected from 207, 268, 317, 360, 661, 697, 779, 876, and 938, as numbered according to SEQ ID NO: 6.

[0581] In some embodiments, an antigenic portion of a Piasmocfium P113 polypeptide in a P113 construct comprises or consists of an amino acid sequence that is at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 135 and comprises one or more substitutions selected from: N207Q, N268Q, N317Q, N360Q, N661Q, N697Q, N779Q, N876Q, and N938Q.

[0582] In some embodiments, an antigenic portion of a Plasmodium P113 polypeptide In a P113 construct comprises or consists of an amino acid sequence of SEQ ID NO: 136.

[0583] In some embodiments, a Plasmodium polypeptide construct described herein further includes one or more linkers. In some embodiments, a Plasmodium polypeptide construct described herein further indudes one or more linkers between one or more regions and/or domains. In some embodiments, a inker comprises or consists of an amino acid sequence according to SEQ ID NO: 86. In some embodiments, a linker comprises or consists of an amino add sequence according to SEQ ID NO: 138.

[0584] In some embodiments, a Plasmodium polypeptide construct described herein further comprises one or more transmembrane regions. In some embodiments, a transmembrane region is a heterologous transmembrane region. In some embodiments, a transmembrane region comprises or consists of a viral transmembrane region. In some embodiments, a transmembrane region comprises or consists of an HSV transmembrane region. In some embodiments, a transmembrane region comprises or consists of an HSV gD transmembrane region. In some embodiments, a transmembrane region comprises or consists of an amino add sequence according to SEQ ID NO: 75.

[0585] In some embodiments, a Plasmodium polypeptide construct can have the following structure:

P113 construct; sec - P113 construct;

Viralsec - Pl 13 construct,

HSV-1gDsec - Pl 13 construct;

P113 construct - TMD; sec - P113 construct - TMD;

Viralsec. - P113 construct - TMD;

HSV-1gDsec. - P113 construct - TMD;

P113 construct - HSV-1TMD; sec - P113 construct - HSV-1TMD;

Viralsec - P113 construct - HSV-1TMD;

HSV-1gDsec. - P113 construct - HSV -1TMD;

P113 construct - Linker - TMD; sec - P113 construct - Linker - TMD;

Viralsec. - P113 construct - Linker - TMD;

HSV-1gDsec. - P113 construct - Linker - TMD; P113 construct - Linker - HSV-1TMD; sec - P113 construct - Linker - HSV-1TMD;

Viralsec - P113 construct - Linker - HSV-1TMD; or HSV-1gDsec. - P113 construct - Linker - HSV-1TMD.

4. Ripr constructs

[0586] In some embodiments, the Plasmodium polypeptide construct described herein includes a Ripr polypeptide or antigenic portion thereof from Plasmodium falciparum, preferably from Plasmodium falciparum isolate 3D7. In some embodiments, the Plasmodium polypeptide construct described herein includes an antigenic portion of Ripr Plasmodium falciparum, preferably from Plasmodium falciparum isolate 3D7 (referred to herein as "Ripr construct" or "Ripr domain").

[0587] In some embodiments, a Plasmodium polypeptide construct described herein includes one or more regions or portions of a Ripr, e.g., Plasmodium Ripr, e.g., P. falciparum Ripr (SEQ ID NO: 2), or a variant thereof (e.g., one or more antigenic portions of a Ripr, e.g., Plasmodium Ripr, e.g., P. falciparum Ripr). In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide in a Ripr construct comprises or consists of an amino add sequence according to SEQ ID NO: 178. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide in a Ripr construct comprises or consists of an amino add sequence with at least 85% sequence Identity to an amino acid sequence according to SEQ ID NO: 178. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide in a Ripr construct comprises or consists of an amino acid sequence according to SEQ ID NO: 177. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide in a Ripr construct comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 177. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide In a Ripr construct comprises or consists of an amino add sequence according to SEQ ID NO: 176. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide In a Ripr construct comprises or consists of an amino acid sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 176. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide In a Ripr construct comprises or consists of an amino add sequence according to SEQ ID NO: 175. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide in a Ripr construct comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 175. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide in a Ripr construct comprises or consists of an amino acid sequence according to SEQ ID NO: 174. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide in a Ripr construct comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 174. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide in a Ripr construct comprises or consists of an amino add sequence according to SEQ ID NO: 173. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide in a Ripr construct comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 173. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide in a Ripr construct comprises or consists of an amino add sequence according to SEQ ID NO: 172. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide in a Ripr construct comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 172. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide in a Ripr construct comprises or consists of an amino acid sequence according to SEQ ID NO: 171. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide in a Ripr construct comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 171.

[0588] In some embodiments, a Plasmodium polypeptide construct comprises one or more antigenic portions of Ripr. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide in a Ripr construct comprises 25, 30, 35, 40, or 45 contiguous amino adds of a Ripr polypeptide, or a variant thereof (e.g., a glycosylation variant). [0589] In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide in a Ripr construct comprises one or more N-lkiked glycosylation sites. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide in a Ripr construct comprises one, two, three, four, five, six, seven, eight, nine, ten, eleven, or twelve N-llnked glycosylation sites. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide In a Ripr construct comprises an amino acid substitution at one or more N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation. In some embodiments, an amino add substitution that prevents glycosylation comprises a NX[T/S] to QX[T/S] substitution. In some embodiments, amino add substitution prevents glycosylation comprises a NX[T/S] to NXA substitution. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide in a Ripr construct comprises an amino acid substitution at all of the N-inked glycosylation sites, wherein the amino add substitution prevents glycosylation.

[0590] In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide in a Ripr construct comprises an asparagine at position 627, position 945, position 1002, or a combination thereof, as numbered according to SEQ ID NO: 190. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide in a Ripr construct comprises a glutamine at position 627, position 945, position 1002, or a combination thereof, as numbered according to SEQ ID NO: 190. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide in a Ripr construct comprises a glutamine at position 627, position 945, and position 1002, as numbered according to SEQ ID NO: 190. In some embodiments, the antigenic portion of an antigenic portion of a Plasmodium Ripr polypeptide In a Ripr construct comprises or consists of an amino add sequence according to SEQ ID NO: 173.

[0591] In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide in a Ripr construct comprises an asparagine at position 84, position 125, position 209, position 284, position 315, position 461, position 479, position 487, position 507, position 627, position 945, position 1002, or a combination thereof, as numbered according to SEQ ID NO: 171. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide in a Ripr construct comprises a glutamine at position 84, position 125, position 209, position 284, position 315, position 461, position 479, position 487, position 507, position 627, position 945, position 1002, or a combination thereof, as numbered according to SEQ ID NO: 171. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide in a Ripr construct comprises a glutamine at position 84, position 125, position 209, position 284, position 315, position 461, position 479, position 487, position 507, position 627, position 945, and position 1002, as numbered according to SEQ ID NO: 171. In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide in a Ripr construct comprises or consists of SEQ ID NO: 172.

[0592] In some embodiments, an antigenic portion of a Plasmodium Ripr polypeptide in a Ripr construct comprises a PMX cleavage site. In some embodiments, a PMX cleavage site of a Ripr antigenic portion in a Ripr construct comprises a sequence of GNISMLEIQNEE (SEQ ID NO: 37). In some embodiments, a Plasmodium Ripr antigenic portion in a Ripr construct does not comprise a PMX cleavage site. In some embodiments, a PMX deavage site of a Ripr antigenic portion in a Ripr construct comprises or consists of a setpence of SMLE (SEQ ID NO: 191). [0593] In some embodiments, a Plasmodium polypeptide construct described herein further includes one or more linkers. In some embodiments, a Plasmodium polypeptide construct described herein further includes one or more linkers between one or more regions and/or domains. In some embodiments, a linker comprises or consists of an amino acid sequence according to SEQ ID NO: 86. In some embodiments, a linker comprises or consists of an amino add sequence according to SEQ ID NO: 138.

[0594] In some embodiments, a Plasmodium polypeptide construct described herein further includes one or more secretory signals. In some embodiments, a secretory signal is a Plasmodium Ripr secretory signal (Riprsec). In some embodiments, a secretory signal comprises or consists of an amino acid sequence according to SEQ ID NO: 186. In some embodiments, a secretory signal is a viral secretory signal. In some embodiments, a secretory signal is an HSV glycoprotein D (gD) secretory signal. In some embodiments, an HSV glycoprotein D (gD) secretory signal comprises or consists of an amino add sequence according to SEQ ID NO: 42.

[0595] In some embodiments, a Plasmodium polypeptide construct described herein further comprises one or more transmembrane regions. In some embodiments, a transmembrane region is a heterologous transmembrane region. In some embodiments, a transmembrane region comprises or consists of a viral transmembrane region. In some embodiments, a transmembrane region comprises or consists of an HSV transmembrane region. In some embodiments, a transmembrane region comprises or consists of an HSV gD transmembrane region. In some embodiments, a transmembrane region comprises or consists of an amino add sequence according to SEQ ID NO: 75.

[0596] In some embodiments, a Plasmodium polypeptide construct described herein further comprises one or more tags. In some embedments, a tag Is a HIBit tag. In some embodiments, a tag comprises or consists of a sequence according to SEQ ID NO: 139.

[0597] In some embodiments, a Plasmodium polypeptide construct can have the following structure:

Ripr construct sec - Ripr construct

Viralsec - Ripr construct

HSV-1gDsec - Ripr construct

Riprsec - Ripr construct

Ripr construct - TMD sec - Ripr construct - TMD

Viralsec - Ripr construct - TMD

HSV-1gDsec. - Ripr construct - TMD

Riprsec - Ripr construct - TMD

Ripr construct - HSV -1TMD sec - Ripr construct - HSV -1TMD

Viralsec - Ripr construct - HSV -1TMD

HSV-1gDsec - Ripr construct - HSV -1TMD

Riprsec - Ripr construct - HSV -1TMD

Ripr construct - muttimerization sec - Ripr construct - muttimerization

Viralsec - Ripr construct - miritimerization NQFCDGEQETCTCKTSLLPSAKNNCEYNDLCTVLNCPEQSTCEQIGNGKKAECKCENG KYYHNNKCYTKNDLELAIKIEPHKKEKFYKNNLYQGKALKPEYIFMQCENGFSIEVINAY VSCYRVSFNLNKLKYVTESLKKMCDGKTKCAYGNTIDPIDDLNHHNICNNFNTIFKYDYL CVFNNQQITSDKNSHLHSNIPSLYQSSILPDIQKSKFHLISRNSRTMQYPHNQISMLEIQ NEISSHNSNQFSTDPHTNSNNINNMNIKKVEIFRSRFSSKLQCQGGKINIDKAILKGGEG CNDLLLTNSLKSYCNDLSECDIGLIYHFD1YCINDQYLFVSYSCSNLCNKCHQNSTCYGN RFNYDCFCDNPYISKYGNKLCERPNDCESVLCSQNQVCQILPNDKLICQCEEGYKNVKG KCVPDNKCDLSCPSNKVCVIENGKQTCKCSERFVLENGVCICANDYKMEDGINCIAKNK CKRKEYENICTNPNEMCAYNEETDIVKCECKEHYYRSSRGECILNDYCKDINCKENEECS IVNFKPECVCKENLKKNNKGECIYENSCLINEGNCPKDSKCIYREYKPHECVCNKQGHVA VNGKCVLEDKCVHNKKCSENSICVNVMNKEPICVCTYNYYKKDGVCLIQNPCLKDNGG CSRNSECTFKYSKIQCTCKENYKNKDDSCVPNTNEYDESFTFQYNDDASIILGACGMIEF SYIYNQIIWKIQNSKESYVFYYDYPTAGNIEVQIKNEIFHniYLKKKIGNSVIYDDFQVD HQTCIYENVFYYSNQNGGSGGGLIAGAVGGSLIAALVICGIVYWMRRHTQKAPKRIRLP HIR

204 57 MGGAAARLGAVILFWIVGLHGVRGLEIQNBSSHNSNQFSTDPHTNSNNINNMNIKKV EIFRSRFSSKLQCQGGKINIDKAILKGGEGCNDLLLTNSLKSYCNDLSECDIGLIYHFDTY CINDQYLFVSYSCSNLCNKCHQNSTCYGNRFNYDCFCDNPYISKYGNKLCERPNDCESV LCSQNQVCQILPNDKUCQCEEGYKNVKGKCVPDNKCDLSCPSNKVCVIENGKQTCKCS ERFVLENGVCICANDYKMEDGINCIAKNKCKRKEYENICTMPNEMCAYNEETDIVKCEC KEHYYRSSRGECILNDYCKDINCKENEECSIVNFKPECVCKENLKKNNKGECIYENSCLI NEGNCPKDSKCIYREYKPHECVCNKQGHVAVNGKCVLEDKCVHNKKCSENSICVNVMN KEPICVCTYNYYKKDGVCLIQNPCLKDNGGCSRNSECTFKYSKIQCTCKENYKNKDDSC VPNTNEYDESFTFQYNDDASIILGACGMIEFSYIYNQIIWKIQNSKESYVFYYDYPTAGN IEVQIKNEIFHTIIYLKKKIGNSVIYDDFQVDHQTCIYENVFYYSNQNGGSGGGLIAGAV GGSLLAALVICGIVYWMRRHTQKAPKRIRLPHIR

205 58 MGGAAARLGAVILFVVIVGLHGVRGDLSCPSNKVCVIENGKQTCKCSERFVLENGVCICA

NDYKMEDGINCIAKNKCKRKEYENICTNPNEMCAYNEETDIVKCECKEHYYRSSRGECI

LNDYCKDINCKENEECSIVNFKPECVCKENLKKNNKGECIYENSCLINEGNCPKDSKCIY

REYKPHECVCNKQGHVAVNGKCVLEDKCVHNKKCSENSICVNVMNKEPICVCTYNYYK

KDGVCUQGGSGGGLIAGAVGGSLLAALVICGIVYWMRRHTQKAPKRIRLPHIR

206 59 MGGAAARLGAVILFWIVGLHGVRGKNKCKRKEYENICTNPNEMCAYNEETDIVKCECK EHYYRSSRGECILNDYCKDINCKENEECSIVNFKPECVCKENLKKNNKGECIYENSCLIN EGNCPKDSKCIYREYKPHECVCNKQGHVAVNGKCVLEDGGSGGGLIAGAVGGSLLAAL VICGIVYWMRRHTQKAPKRIRLPHIR

207 60 MGGAAARLGAVILFWIVGLHGVRGKNKCKRKEYENICTNPNEMCAYNEETDIVKCECK EHYYRSSRGECILNDYCKDINCKENEECSIVNFKPECVCKENLKKNNKGECIYEGGSGGG LIAGAVGGSLLAALVICG1VYWMRRHTQKAPKRIRLPHIR

208 61 MGGAAARLGAV1LFWIVGLHGVRGDYCKDINCKENEECSIVNFKPECVCKENLKKNNK

GECIYENSCLINEGNCPKDSKCIYREYKPHECVCNKQGHVAVNGKCVLEDGGSGGGLIA

GAVGGSLLAALVICGIVYWMRRHTQKAPKRIRLPHIR

209 62 MGGAAARLGAVILFWrVGLHGVRGDYCKDINCKENEECSIVNFKPECVCKENLKKNNK

GECIYEGGSGGGUAGAVGGSLLAALVICGIVYWMRRHTQKAPKRIRLPHIR

[0599] In some embodiments, a Ripr construct described herein has an amino add sequence provided in Table 13. In some embodiments, a Ripr construct comprises an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino acid sequence of Table 13. [0600] In some embodiments, a Ripr construct comprises or consists of an amino add sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino add sequence of SEQ ID NO: 145. In some embodiments, a Ripr construct comprises or consists of an amino add sequence of SEQ ID NO: 145.

[0601] In some embodiments, a Ripr construct comprises or consists of an amino add sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino add sequence of SEQ ID NO: 146. In some embodiments, a Ripr construct comprises or consists of an amino add sequence of SEQ ID NO: 146.

[0602] In some embodiments, a Ripr construct comprises or consists of an amino add sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino add sequence of SEQ ID NO: 147. In some embodiments, a Ripr construct comprises or consists of an amino acid sequence of SEQ ID NO: 147.

[0603] In some embodiments, a Ripr construct comprises or consists of an amino add sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino add sequence of SEQ ID NO: 148. In some embodiments, a Ripr construct comprises or consists of an amino add sequence of SEQ ID NO: 148.

[0604] In some embodiments, a Ripr construct comprises or consists of an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino add sequence of SEQ ID NO: 149. In some embodiments, a Ripr construct comprises or consists of an amino acid sequence of SEQ ID NO: 149.

[0605] In some embodiments, a Ripr construct comprises or consists of an amino add sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% Identity to an amino add sequence of SEQ ID NO: 150. In some embodiments, a Ripr construct comprises or consists of an amino add sequence of SEQ ID NO: 150.

[0606] In some embodiments, a Ripr construct comprises or consists of an amino add sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% Identity to an amino add sequence of SEQ ID NO: 151. In some embodiments, a Ripr construct comprises or consists of an amino acid sequence of SEQ ID NO: 151.

[0607] In some embodiments, a Ripr construct comprises or consists of an amino add sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% Identity to an amino add sequence of SEQ ID NO: 152. In some embodiments, a Ripr construct comprises or consists of an amino add sequence of SEQ ID NO: 152.

[0608] In some embodiments, a Ripr construct comprises or consists of an amino add sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% Identity to an amino add sequence of SEQ ID NO: 153. In some embodiments, a Ripr construct comprises or consists of an amho acid sequence of SEQ ID NO: 153.

[0609] In some embodiments, a Ripr construct comprises or consists of an amino add sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino add sequence of SEQ ID NO: 154. In some embodiments, a Ripr construct comprises or consists of an amino add sequence of SEQ ID NO: 154.

[0610] In some embodiments, a Ripr construct comprises or consists of an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% Identity to an amino add sequence of SEQ ID NO: 155. In some embodiments, a Ripr construct comprises or consists of an amino add sequence of SEQ ID NO: 155.

[0611] In some embodiments, a Ripr construct comprises or consists of an amino add sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino add sequence of SEQ ID NO: 156. In some embodiments, a Ripr construct comprises or consists of an amino add sequence of SEQ ID NO: 156.

[0612] In some embodiments, a Ripr construct comprises or consists of an amino add sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino add sequence of SEQ ID NO: 157. In some embodiments, a Ripr construct comprises or consists of an amino add sequence of SEQ ID NO: 157.

[0624] In some embodiments, a Ripr construct described herein has a nudek add sequence provided in Table

14. In some embodiments, a Ripr construct comprises a nucleic add sequence having at least 99%, 98%, 97%,

96%, 95%, 90%, 85%, or 80% identity to nudek add sequence of Table 14.

Table 15: RNA Sequences Corresponding to Exemplary Ripr RNA Constructs as Described Herein [0625] In some embodiments, a Ripr construct described herein has a nudeic add sequence provided in Table 15. In some embodiments, a Ripr construct comprises a nudeic add sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to a nucleic acid sequence of Table 15.

5. TRAMP constructs

[0626] In some embodiments, the Plasmodium polypeptide construct described herein includes a TRAMP polypeptide or antigenic portion thereof from Plasmodium falciparum, preferably from Plasmodium falciparum isolate 3D7. In some embodiments, the Plasmodium polypeptide construct described herein includes an antigenic portion of TRAMP Plasmodium falciparum, preferably from Plasmodium falciparum isolate 3D7 (referred to herein as "TRAMP construct" or "TRAMP domain").

[0627] In some embodiments, a Plasmodium polypeptide construct described herein includes one or more regions or portions of a TRAMP, e.g., Hasmodium TRAMP, e.g., P. faldparumTRAW? (SEQ ID NO: 4), or a variant thereof (e.g., one or more antigenic portions of a TRAMP, e.g., Plasmodium TRAMP, e.g., P. faidparum TRAMP). In some embodiments, an antigenic portion of a Plasmodium TRAMP polypeptide in a TRAMP construct comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 179. In some embodiments, an antigenic portion of a Hasmodium TRAMP polypeptide in a TRAMP construct comprises or consists of an amino acid sequence according to SEQ ID NO: 179.

[0628] In some embodiments, a Plasmodium TRAMP polypeptide or antigenic portion thereof comprises 25, 30, 35, 40, or 45 contiguous amino adds of a TRAMP polypeptide, or a variant thereof (e.g., a glycosylation variant). In some embodiments, a Plasmodium polypeptide construct comprises one or more antigenic portions of TRAMP.

[0629] In some embodiments, an antigenic portion of a Plasmodium TRAMP polypeptide in a TRAMP construct comprises one or more N-linked glycosylation sites. In some embodiments, an antigenic portion of a Plasmodium TRAMP polypeptide In a TRAMP construct comprises one, two, three, four, five, six, seven, or eight N-llnked glycosylation sites. In some embodiments, an antigenic portion of a Hasmodium TRAMP polypeptide In a TRAMP construct comprises an amino add substitution at one or more N-llnked glycosylation sites, wherein the amino add substitution prevents glycosylation. In some embodiments, an amino acid substitution that prevents glycosylation comprises a NX[T/S] to QX[T/S] substitution. In some embodiments, amino acid substitution prevents glycosylation comprises a NX[T/S] to NXA substitution. In some embodiments, an antigenic portion of a PlasmodiumTWdAP polypeptide in a TRAMP construct comprises an amino acid substitution at all of the N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation.

[0630] In some embodiments, an antigenic portion of a PlasmodiumTRMAP polypeptide in a TRAMP construct comprises an asparagine at position 108, position 154, position 161, or a combination thereof, as numbered according to SEQ ID NO: 179. In some embodiments, an antigenic portion of a Plasmodium TRAMP polypeptide in a TRAMP construct comprises a glutamine at position 108, position 154, position 161, or a combination thereof, as numbered according to SEQ ID NO: 179. In some embodiments, an antigenic portion of a Plasmodium TRAMP polypeptide in a TRAMP construct comprises a glutamine at position 108, position 154, and position 161, as numbered according to SEQ ID NO: 179. In some embodiments, an antigenic portion of a Plasmodium TRAMP polypeptide in a TRAMP construct comprises or consists of an amino acid sequence according to SEQ ID NO: 181. [0631] In some embodiments, an antigenic portion of a Plasmodium TRAMP polypeptide in a TRAMP construct comprises an asparagine at position 71, position 108, position 114, position 129, position 154, position 161, position 212, position 264, or a combination thereof, as numbered according to SEQ ID NO: 179. In some embodiments, an antigenic portion of a Plasmodium TRAMP polypeptide in a TRAMP construct comprises a glutamine at position 71, position 108, position 114, position 129, position 154, position 161, position 212, position 264, or a combination thereof, as numbered according to SEQ ID NO: 179. In some embodiments, an antigenic portion of a Plasmodium TRAMP polypeptide in a TRAMP construct comprises a glutamine at position 71, position 108, position 114, position 129, position 154, position 161, position 212, and position 264, as numbered according to SEQ ID NO: 179. In some embodiments, an antigenic portion of a Plasmodium TRAMP polypeptide in a TRAMP construct comprises or consists of an amino add sequence according to SEQ ID NO: 180.

[0632] In some embodiments, an antigenic portion of a Plasmodium TRAMP polypeptide in a TRAMP construct comprises a PMX deavage site. In some embodiments, a PMX cleavage site of a TRAMP polypeptide or antigenic portion thereof comprises or consists of a sequence of HFLQ (SEQ ID NO: 192). In some embodiments, an antigenic portion of a Plasmodium TRAMP polypeptide in a TRAMP construct comprises a SUB2 deavage site. In some embodiments, a SUB2 deavage site of a TRAMP polypeptide or antigenic portion thereof comprises or consists of ILMIILPIYLVISIITILYHIFY (SEQ ID NO: 193).

[0633] In some embodiments, a Plasmodium polypeptide construct described herein further Includes one or more linkers. In some embodiments, a Plasmodium polypeptide construct described herein further includes one or more linkers between one or more regions and/or domains. In some embodiments, a inker comprises or consists of an amino acid sequence according to SEQ ID NO: 86. In some embodiments, a linker comprises or consists of an amino add sequence according to SEQ ID NO: 138.

[0634] In some embodiments, a Plasmodium polypeptide construct described herein further Includes one or more secretory signals. In some embodiments, a secretory signal Is a Plasmodium TRAMP secretory signal (TRAMPsec). In some embodiments, a secretory signal comprises or consists of an amino acid sequence according to SEQ ID NO: 219. In some embodiments, a secretory signal is a viral secretory signal. In some embodiments, a secretory signal is an HSV glycoprotein D (gD) secretory signal. In some embodiments, an HSV glycoprotein D (gD) secretory signal comprises or consists of an amino add sequence according to SEQ ID NO: 42.

[0635] In some embodiments, a Plasmodium polypeptide construct described herein further comprises one or more transmembrane regions. In some embodiments, a transmembrane region Is a heterologous transmembrane region. In some embodiments, a transmembrane region comprises or consists of a viral transmembrane region. In some embodiments, a transmembrane region comprises or consists of an HSV transmembrane region. In some embodiments, a transmembrane region comprises or consists of an HSV gD transmembrane region. In some embodiments, a transmembrane region comprises or consists of an amino add sequence according to SEQ ID NO: 75.

[0636] In some embodiments, a Plasmodium polypeptide construct described herein further comprises one or more tags. In some embodiments, a tag is a HiBit tag. In some embodiments, a tag comprises or consists of a sequence according to SEQ ID NO: 139.

[0637] In some embodiments, a Plasmodium polypeptide construct can have the following structure:

TRAMP construct sec - TRAMP construct

Vrateec. - TRAMP construct

HSV-lgDsec - TRAMP construct

[0639] In some embodiments, a TRAMP construct described herein has an amino add sequence provided in Table 16. In some embodiments, a TRAMP construct comprises an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino add sequence of Table 16.

[0640] In some embodiments, a TRAMP construct comprises or consists of an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino add sequence of SEQ ID NO: 159. In some embodiments, a TRAMP construct comprises or consists of an amino add sequence of SEQ ID NO: 159.

[0641] In some embodiments, a TRAMP construct comprises or consists of an amino add sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino add sequence of SEQ ID NO: 160. In some embodiments, a TRAMP construct comprises or consists of an amino add sequence of SEQ ID NO: 160.

[0642] In some embodiments, a TRAMP construct comprises or consists of an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino add sequence of SEQ ID NO: 161. In some embodiments, a TRAMP construct comprises or consists of an amino add sequence of SEQ ID NO: 161.

Table 17: DNA Sequences Corresponding to Exemplary TRAMP RNA Constructs as Described Herein

[0643] In some embodiments, a TRAMP construct described herein has a nucleic add sequence provided In

Table 17. In some embodiments, a TRAMP construct comprises a nudek add sequence having at least 99%, 98%,

97%, 96%, 95%, 90%, 85%, or 80% Identity to a nudek acid sequence of Table 17.

Table 18: RNA Sequences Corresponding to Exemplary TRAMP RNA Constructs as Described Herein 6. CSS constructs

[0645] In some embodiments, the Plasmodium polypeptide construct described herein includes a CSS polypeptide or antigenic portion thereof from Piasmodium falciparum, preferably from Plasmodium falciparum isolate 3D7. In some embodiments, the Piasmodium polypeptide construct described herein includes an antigenic portion of CSS Piasmodium faldparum, preferably from Plasmodium faldparum isolate 3D7 (referred to herein as “CSS construct” or "CSS domain").

[0646] In some embodiments, a Plasmodium polypeptide construct described herein includes one or more regions or portions of a CSS, e.g., Piasmodium CSS, e.g., P. falciparum CSS (SEQ ID NO: 5), or a variant thereof (e.g., one or more antigenic portions of a CSS, e.g., Plasmodium CSS, e.g., P. falciparum CSS). In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide In a CSS construct comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 194. In some embodiments, an antigenic portion of a Piasmodium CSS polypeptide In a CSS construct comprises or consists of an amino add sequence according to SEQ ID NO: 194. In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide in a CSS construct comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 195. In some embodiments, an antigenic portion of a Piasmodium CSS polypeptide in a CSS construct comprises or consists of an amino acid sequence according to SEQ ID NO: 195. In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide In a CSS construct comprises or consists of an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 182. In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide in a CSS construct comprises or consists of an amino add sequence according to SEQ ID NO: 182. [0647] In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide in a CSS construct comprises 25, 30, 35, 40, or 45 contiguous amino acids of a CSS polypeptide, or a variant thereof (e.g., a glycosylation variant). In some embodiments, a Plasmodium polypeptide construct comprises one or more antigenic portions of CSS.

[0648] In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide In a CSS construct comprises one or more N-linked glycosylation sites. In some embodiments, an antigenic portion of a Piasmodium CSS polypeptide in a CSS construct comprises one, two, three, four, five, or six N-linked glycosylation sites. In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide in a CSS construct comprises an amino acid substitution at one or more N-linked glycosylation sites, where the amino acid substitution prevents glycosylation. In some embodiments, an amino add substitution that prevents glycosylation comprises a NX[T/S] to QX[T/S] substitution. In some embodiments, amino add substitution prevents glycosylation comprises a NX[T/S] to NXA substitution. In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide In a CSS construct comprises an amino acid substitution at all of the N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation.

[0649] In some embodiments, an antigenic portion of a Piasmodium CSS polypeptide in a CSS construct comprises an asparagine at position 54, position 68, or a combination thereof, as numbered according to SEQ ID NO: 194. In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide in a CSS construct comprises a glutamine at position 54, position 68, or a combination thereof, as numbered according to SEQ ID NO: 194. In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide in a CSS construct comprises a glutamine at position 54, and position 68, as numbered according to SEQ ID NO: 194. In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide in a CSS construct comprises or consists of an amino acid sequence according to SEQ ID NO: 184.

[0650] In some embodiments, an antigenic portion of a Piasmodium CSS polypeptide in a CSS construct comprises an asparagine at position 172, position 214, position 241, position 263, or a combination thereof, as numbered according to SEQ ID NO: 195. In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide in a CSS construct comprises a glutamine at position 172, position 214, position 241, position 263, or a combination thereof, as numbered according to SEQ ID NO: 195. In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide in a CSS construct comprises a glutamine at position 172, position 214, position 241, and position 263, as numbered according to SEQ ID NO: 195. In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide In a CSS construct comprises or consists of an amino add sequence according to SEQ ID NO: 185.

[0651] In some embodiments, an antigenic portion of a Piasmodium CSS polypeptide in a CSS construct comprises an asparagine at position 54, position 68, position 172, position 214, position 241, position 263, or a combination thereof, as numbered according to SEQ ID NO: 182. In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide in a CSS construct comprises a glutamine at position 54, position 68, position 172, position 214, position 241, position 263, or a combination thereof, as numbered according to SEQ ID NO: 182. In some embodiments, an antigenic portion of a Plasmodium CSS polypeptide In a CSS construct comprises a glutamine at position 54, position 68, position 172, position 214, position 241, and position 263, as numbered according to SEQ JD NO: 182. In some embodiments, an antigenic portion of a Piasmodium CSS polypeptide in a CSS construct comprises or consists of an amino acid sequence according to SEQ ID NO: 183.

[0652] In some embodiments, a Plasmodium polypeptide construct described herein further includes one or more linkers. In some embodiments, a Plasmodium polypeptide construct described herein further Includes one or more linkers between one or more regions and/or domains. In some embodiments, a linker comprises or consists of an amino acid sequence according to SEQ ID NO: 86. In some embodiments, a linker comprises or consists of an amino add sequence according to SEQ ID NO: 138.

[0653] In some embodiments, a Plasmodium polypeptide construct described herein further Includes one or more secretory signals. In some embodiments, a secretory signal Is a Plasmodium CSS secretory signal (CSSsec). In some embodiments, a secretory signal comprises or consists of an amino acid sequence according to SEQ ID NO: 188. In some embodiments, a secretory signal is a viral secretory signal. In some embodiments, a secretory signal is an HSV glycoprotein D (gD) secretory signal. In some embodiments, an HSV glycoprotein D (gD) secretory signal comprises or consists of an amino add sequence according to SEQ ID NO: 42.

[0654] In some embodiments, a Plasmodium polypeptide construct described herein further comprises one or more transmembrane regions. In some embodiments, a transmembrane region is a heterologous transmembrane region. In some embodiments, a transmembrane region comprises or consists of a viral transmembrane region. In some embodiments, a transmembrane region comprises or consists of an HSV transmembrane region. In some embodiments, a transmembrane region comprises or consists of an HSV gD transmembrane region. In some embodiments, a transmembrane region comprises or consists of an amino add sequence according to SEQ ID NO: 75. [0655] In some embodiments, a Plasmodium polypeptide construct described herein further comprises one or more tags. In some emtxxfiments, a tag is a HiBit tag. In some embodiments, a tag comprises or consists of a sequence according to SEQ ID NO: 139.

[0656] In some embodiments, a Plasmodium polypeptide construct can have the following structure:

SEQ RNA Construct Sequence (Amino Add)

ID

NO:

162 44 MIKVLLAVLFILIKLENIIGQDEKSVKNISVCDFTDKLNFLPLEKTKILCELKPQYGEDIKII ANKEYEINCMNNSKVFCPLKDTFINNTNIKLYSPKLHFEIKDITHKGKNAALYYLKIDEEA SDIFFSCSIKPKQVSGLLEGEVRVNLKKHINEEYSIFNEEEDVHVCDFSKGNLDITPSAGF YLKNSRNVSClYRVIPNKLFLIKLPKLDIVrEKLLPSIVNCLSEFSFINFTLKHVQEGDNYIS FNVIFGEFKKHFNLTCSLDLSDFQQEPCNLGKTANITFIFSK

163 45 MIKVLLAVLFILIKLENIIGQDEKSVKNISVCDFTDKLNFLPLEKTKILCELKPQYGEDIKII ANKEYEINCMQNSKVFCPLKDTHQNTNIKLYSPKLHFEIKDITHKGKNAALYYLKIDEEA SDIFFSCSIKPKQVSGLLEGEVRVNLKKHINEEYSIFNEEEDVHVCDFSKGNLDITPSAGF YLKNSRQVSCIYRVIPNKLFLIKLPKLDIVTEKLLPSIVNCLSEFSFIQFTLKHVQEGDNYIS FNVIFGEFKKHFQLTCSLDLSDFQQEPCNLGKTAQITFIFSK

164 46 MGGAAARLGAVILFWIVGLHGVRGQDEKSVKNISVCDFTDKLNFLPLEKTKILCELKPQ YGEDIKIIANKEYEINCMQNSKVFCPLKDTFIQNTNIKLYSPKLHFBKDITHKGKNAALY YLKIDEEASDIFFSCSIKPKQVSGLLEGEVRVNLKKHINEEYSIFNEEEDVHVCDFSKGNL DITPSAGFYLKNSRQVSCIYRVIPNKLFLIKLPKLDIVTEKLLPSIVNCLSEFSFIQFTLKHV QEGDNYISFNVIFGEFKKHFQLTCSLDLSDFQQEPCNLGKTAQITFIFSK

165 47 MIKVLLAVLFILIKLENIIGQDEKSVKNISVCDFTDKLNFLPLEKTKILCELKPQYGEDIKII ANKEYEINCMQNSKVFCPLKDTnQNTNIKLYSPKLHFEIKDITHKGKNAALYYLKIDEEA SDIFFSCSIKPKQVSGLLEGEVRVNLKKHINEEYSIFNEEEDVHVCDFSKGNLDITPSAGF YLKNSRQVSCIYRVIPNKLFUKLPKLDIVTEKLLPSIVNCLSEFSFIQFTLKHVQEGDNYIS FNVIFGEFKKHFQLTCSLDLSDFQQEPCNLGKTAQITFIFSKGLIAGAVGGSLLAALVICG JVYWMRRHTQKAPKRIRLPHIR

166 48 MGGAAARLGAVILFWIVGLHGVRGQDEKSVKNISVCDFTDKLNFLPLEKTKILCELKPQ YGEDIKIIANKEYEINCMQNSKVFCPLKDTHQNTNIKLYSPKLHFEIKDITHKGKNAALY YLKIDEEASDIFFSCSIKPKQVSGLLEGEVRVNLKKHINEEYSIFNEEEDVHVCDFSKGNL DITPSAGFYLKNSRQVSCIYRVIPNKLFLIKLPKLDIVrEKLLPSIVNCLSEFSFIQFTLKHV QEGDNYISFNVIFGEFKKHFQLTCSLDLSDFQQEPCNLGKTAQITHFSKGLIAGAVGGS

LLAALVICGIVYWMRRHTQKAPKRIRLPHIR

167 49 MGGAAARLGAVILFWIVGLHGVRGQDEKSVKNISVCDFTDKLNFLPLEKTKILCELKPQ YGEDIKIIANKEYEINCMQNSKVFSPLKDTFIQNTNIKLYSPKLHFEIKDTTHKGKNAALY YLKIDEEASDIFFSCSIKPKQVSGLLEGEVRVNLKKGLIAGAVGGSLLAALVICGIVYWMR RHTDKAPKRIRLPHIR

168 50 MGGAAARLGAVILFWIYGLHGVRGHINEEYSIFNEEEDVHVCDFSKGNLDITPSAGFYL KNSRQVSCIYRVIPNKLFLIKLPKLDIVTEKLLPSIVNCLSEFSFIQFTLKHVQEGDNYISF NVIFGEFKKHFQLTCSLDLSDFQQEPSNLGKTAQITFIFSKGLIAGAVGGSLLAALVICGI VYWMRRHTQKAPKRIRLPHIR

210 63 MlKVLLAVLFILIKLENIIGQDEKSVKNISVCDFTDKLNFLPLEKTKILCELKPQYGEDIKII

ANKEYEINCMQNSKVFCPLKDTFIQNTNIKLYSPKLHFEIKDITHKGKNAALYYLKIDEEA

SDIFFSCSIKPKQVSGLLEGEVRVNLKKHINEEYSIFNEEEDVHVCDFSKGNLDITPSAGF YLKNSRQVSCIYRVIPNKLFLIKLPKLDIVTEKLLPSIVNCLSEFSFIQFTLKHVQEGDNYIS FNVIFGEFKKHFQLTCSLDLSDFQQEPCNLGKTAQITHFSKGGSGGGLIAGAVGGSLLA ALVICGIVYWMRRHTQKAPKRIRLPHIR _

211 64 MGGAAARLGAVILFWIVGLHGVRGQDEKSVKNISVCDFTDKLNFLPLEKTKILCELKPQ YGEDIKIIANKEYEINCMQNSKVFCPLKDTFIQNTTNIKLYSPKLHFBKDITHKGKNAALY YLKIDEEASDIFFSCSIKPKQVSGLLEGEVRVNLKKHINEEYSIFNEEEDVHVCDFSKGNL DITPSAGFYLKNSRQVSCIYRVIPNKLFUKLPKLDIVTEKLLPSIVNCLSEFSFIQFTLKHV QEGDNYISFNVIFGEFKKHFQLTCSLDLSDFQQEPCNLGKTAQITFIFSKGGSGGGLIAG AVGGSLLAALVICGIVYWMRRHTQKAPKRIRLPHIR _

212 65 MGGAAARLGAVILFWIVGLHGVRGQDEKSVKNISVCDFTDKLNFLPLEICrXILCELKPQ YGEDIKIIANKEYEINCMQNSKVFSPLKDTFIQNTNIKLYSPKLHFEIKDITHKGKNAALY YLKIDEEASDIFFSCSIKPKQVSGLLEGEVRVNLKKGGSGGGUAGAVGGSLLAALVICGI VYWMRRHTQKAPKRIRLPHIR _

213 66 MGGAAARLGAVlLFWIVGLHGVRGHINEEYSIFNEEEDVHVCDFSKGNLDnPSAGFYL KNSRQVSCIYRVIPNKLFLIKLPKLDIVTEKLLPSIVNCLSEFSFIQFTLKHVQEGDNYISF NVIFGEFKKHFQLTCSLDLSDFQQEPSNLGKTAQrTFIFSKGGSGGGLIAGAVGGSLLAA LVICGIVYWMRRHTQKAPKRIRLPHIR _

336 132 MIKVLLAVLFIUKUENIIGQDEKSVKNICVCDFTDKLNFLPLEKTKILCELKPQYGEDIKn ANKEYEINCMNNSKVFCPLKDTFINNTNIKLYSPKLHFBKDITHKGKNAALYYLKIDEEA SDIFFSCSIKPKQVSGLLEGEVRVNLKKHINEEYSIFNEEEDVHVCDFSKGNLDITPSAGF YLKNSRNVSCIYRVIPNKLFLIKLPKLDIVTEKLLPSIVNCLSEFSFINFTLKHVQEGDNYIS FNVIFGEFKKHFNLTCSLDLSDFQQEPCNLGICTANrTHFSK

[0658] In some embodiments, a CSS construct described herein has an amino acid sequence provided in Table 19. In some embodiments, a CSS construct comprises an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino add sequence of Table 19.

[0659] In some embodiments, a CSS construct comprises or consists of an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% Identity to an amino acid sequence of SEQ ID NO: 162. In some embodiments, a CSS construct comprises or consists of an amino add sequence of SEQ ID NO: 162.

[0660] In some embodiments, a CSS construct comprises or consists of an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino add sequence of SEQ ID NO: 163. In some embodiments, a CSS construct comprises or consists of an amino add sequence of SEQ ID NO: 163.

[0661] In some embodiments, a CSS construct comprises or consists of an amino add sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino add sequence of SEQ ID NO: 164. In some embodiments, a CSS construct comprises or consists of an amino add sequence of SEQ ID NO: 164.

[0662] In some embodiments, a CSS construct comprises or consists of an amino add sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% Identity to an amino add sequence of SEQ ID NO: 165. In some embodiments, a CSS construct comprises or consists of an amino add sequence of SEQ ID NO: 165.

[0663] In some embodiments, a CSS construct comprises or consists of an amino add sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% Identity to an amino add sequence of SEQ ID NO: 166. In some embodiments, a CSS construct comprises or consists of an amino add sequence of SEQ ID NO: 166.

[0664] In some embodiments, a CSS construct comprises or consists of an amino add sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino add sequence of SEQ ID NO: 167. In some embodiments, a CSS construct comprises or consists of an amino add sequence of SEQ ID NO: 167.

[0665] In some embodiments, a CSS construct comprises or consists of an amino add sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino add sequence of SEQ ID NO: 168. In some embodiments, a CSS construct comprises or consists of an amino add sequence of SEQ ID NO: 168. [0666] In some embodiments, a CSS construct comprises or consists of an amino add sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino add sequence of SEQ ID NO: 210. In some embodiments, a CSS construct comprises or consists of an amino acid sequence of SEQ ID NO: 210.

[0667] In some embodiments, a CSS construct comprises or consists of an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino add sequence of SEQ ID NO: 211. In some embodiments, a CSS construct comprises or consists of an amino add sequence of SEQ ID NO: 211.

[0668] In some embodiments, a CSS construct comprises or consists of an amino add sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino add sequence of SEQ ID NO: 212. In some embodiments, a CSS construct comprises or consists of an amino add sequence of SEQ ID NO: 212.

[0669] In some embodiments, a CSS construct comprises or consists of an amino add sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino add sequence of SEQ ID NO: 213. In some embodiments, a CSS construct comprises or consists of an amino add sequence of SEQ ID NO: 213.

Table 20: DNA Sequences Corresponding to Exemplary CSS RNA Constructs as Described Herein

[0670] In some embodiments, a CSS construct described herein has a nucleic add sequence provided In Table

20. In some embodiments, a CSS construct comprises a nudek add sequence having at least 99%, 98%, 97%,

96%, 95%, 90%, 85%, or 80% identity to a nucleic add sequence of Table 20.

Table 21: RNA Sequences Corresponding to Exemplary CSS RNA Constructs as Described Herein [0671] In some embodiments, a CSS construct described herein has a nucleic add sequence provided in Table 21. In some embodiments, a CSS construct comprises a nudeic acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to a nucleic acid sequence of Table 21.

7. GARP constructs

[0672] In some embodiments, the Plasmodium polypeptide construct described herein includes a GARP polypeptide or antigenic portion thereof from Plasmodium falciparum, preferably from Plasmodium falciparum isolate 3D7. In some embodiments, the Plasmodium polypeptide construct described herein includes an antigenic portion of GARP Plasmodium faiciparum, preferably from Plasmodium falciparum isolate 3D7 (referred to herein as "GARP construct" or "GARP domain").

[0673] In some embodiments, a Plasmodium polypeptide construct described herein Includes one or more regions or portions of a GARP, e.g., Plasmodium GARP, e.g., P. faldparum GARP (SEQ ID NO: 23), or a variant thereof (e.g., one or more antigenic portions of a GARP, e.g., Plasmodium GARP, e.g., P. faldparum GARP). In some embodiments, an antigenic portion of a Plasmodium GARP polypeptide in a GARP construct comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 312. In some embodiments, an antigenic portion of a Plasmodium GARP polypeptide in a GARP construct comprises or consists of an amino acid sequence according to SEQ ID NO: 312. In some embodiments, an antigenic portion of a Plasmodium GARP polypeptide In a GARP construct comprises or consists of an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 313. In some embodiments, an antigenic portion of a Plasmodium GARP polypeptide in a GARP construct comprises or consists of an amino add sequence according to SEQ ID NO: 313. In some embodiments, an antigenic portion of a Plasmodium GARP polypeptide in a GARP construct comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 314. In some embodiments, an antigenic portion of a Plasmodium GARP polypeptide In a GARP construct comprises or consists of an amino add sequence according to SEQ ID NO: 314.

[0674] In some embodiments, an antigenic portion of a Plasmodium GARP polypeptide In a GARP construct comprises 25, 30, 35, 40, or 45 contiguous amino adds of a GARP polypeptide, or a variant thereof (e.g., a glycosylation variant). In some embodiments, a Plasmodium polypeptide construct comprises one or more antigenic portions of GARP.

[0675] In some embodiments, an antigenic portion of a Plasmodium GARP polypeptide in a GARP construct comprises one or more N-linked glycosylation sites. In some embodiments, an antigenic portion of a Plasmodium GARP polypeptide in a GARP construct comprises one, two, three, four, five, or six N-linked glycosylation sites. In some embodiments, an antigenic portion of a Plasmodium GARP polypeptide in a GARP construct comprises an amino add substitution at one or more N-llnked glycosylation sites, where the amino add substitution prevents glycosylation. In some embodiments, an amino add substitution that prevents glycosylation comprises a NX[T/S] to QX[T/S] substitution. In some embodiments, amino acid substitution prevents glycosylation comprises a NX[T/S] to NXA substitution. In some embodiments, an antigenic portion of a Plasmodium GARP polypeptide in a GARP construct comprises an amino acid substitution at all of the N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation.

Table 22: Amino Acid Sequences Encoded by Exemplary GARP RNA Constructs as Described Herein

[0676] In some embodiments, a GARP construct described herein has an amino acid sequence provided in Table 22. In some embodiments, a GARP construct comprises an amino add sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to an amino add sequence of Table 22.

Table 23: DNA Sequences Corresponding to Exemplary GARP RNA Constructs as Described Herein ACCAGACCTCCGAGGAAACAAACAACGAAATCATGGTGCCCCTGCCTTCTCCACTGA CCGACGTGACAACACCTGAGGAACACAAAGAGGGCGAGCACAAAGAGGAAGAACACA AAGAAGGGGAACACAAAGAGGGCGAACACAAAGAAGAAGAACACAAAGAGGAAGAA CATAAGAAAGAGGAACATAAGAGCAAAGAGCACAAGTCCAAGGGCAAGAAGGACAAG GGGAAGAAAGATAAGGGCAAGCACAAGAAGGCCAAGAAAGAGAAAGTCAAGAAGCA

CGTCGTCAAGAACGTGATCGAGGACGAGGACAAGGACGGCGTGGAAATCATCAACCT TGAGGACAAAGAGGCCTGCGAGGAACAGCACATCACCGTGGAAAGCAGACCCCTGTC TCAGCCCCAGTGCAAGCTGATCGATGAGCCTGAGCAGCTGACCCTGATGGACAAGAG CAAGGTCGAGGAAAAGCAGCTGAGCATCCAAGAACAGCTGATCGGCACCATCGGCAG AGTGAACGTGGTGCCCAGACGGGACAACCACAAGAAAAAGATGGCCAAGATCGAGG

AAGCCGAGCTGCAAAAGCAGAAACACGTCGACAAAGAAGAAGATAAGAAAGAGGAAT CCAAAGAGGTTGAGGAAGAAAGCAAAGAGGTCCAAGAGGACGAAGAAGAAGTTGAG GAGGACGAGGAAGAAGAAGAAGAAGAGGAAGAGGAAGAAGAAGAAGAGGAAGAGGA AGAAGAAGAAGAAGAGGACGAGGTCGAGGAAGATGAGGACGACGCCGAAGAAGATG AGGACGACGCTGAGGAAGATGAGGACGACGCCGAGGAAGATGACGACGACGCTGAA

GAAGATGACGACGACGCCGAAGAAGATGATGATGAGGACGAGGACGAAGATGAGGA AGAAGAAGAAGACGAGGAAGAAGAAGAAGAATCCGAGAAGAAGATCAAGCGGAACC TGCGGAAGAACGCCAAGATCGGAGGCTCTGGCGGTGGACTTATTGCCGGCGCTGTT GGAGGATCTCTGCTGGCCGCTCTGGTCATCTGTGGCATCGTCTACTGGATGCGGCG GCACACACAGAAGGCCCCTAAGAGAATCAGACTGCCCCACATCAGATGATGA

[0677] In some embodiments, a GARP construct described herein has a nucleic add sequence provided in

Table 23. In some embodiments, a GARP construct comprises a nucleic add sequence having at least 99%, 98%,

97%, 96%, 95%, 90%, 85%, or 80% Identity to a nudek add sequence of Table 23.

Table 24: RNA Sequences Corresponding to Exemplary GARP RNA Constructs as Described Herein

SEQ RNA Construct Sequence (RNA)

ID

NO:

302 77 AUGGGCGGAGCUGCUGCUAGACUGGGAGCCGUGAUCCUGUUCGUGGUUAUCGUG

GGACUGCAUGGCGUGCGGGGCGACAAGAGCAAAGAGCACAAGUCCAAGGGCAAGAA

GGACAAGGGGAAGAAAGAUAAGGGCAAGCACAAGAAGGCCAAGAAAGAGAAAGUCA

AGAAGCACGUCGUCAAGAACGUGAUCGAGGACGAGGACAAGGACGGCGUGGAAAUC

AUCAACCUUGAGGACAAAGAGGCCUGCGAGGAACAGCACAUCACCGUGGAAAGCAG

ACCCCUGUCUCAGCCCCAGUGCAAGCUGAUCGAUGAGCCUGAGCAGCUGACCCUGA

UGGACAAGAGCAAGGUCGAGGAAAAGAACCUGAGCAUCCAAGAACAGCUGAUCGGC

ACCAUCGGCAGAGUGAACGUGGUGCCCAGACGGGACAACCACAAGAAAAAGAUGGC

CAAGAUCGAGGAAGCCGAGCUGCAAAAGCAGAAACACGUCGACAAAGAAGAAGAUA

AGAAAGAGGAAUCCAAAGAGGUUGAGGAAGAAAGCAAAGAGGUCCAAGAGGACGAA

GAAGAAGUUGAGGAGGACGAGGAAGAAGAAGAAGAAGAGGAAGAGGAAGAAGAAG

AAGAGGAAGAGGAAGAAGAAGAAGAAGAGGACGAGGUCGAGGAAGAUGAGGACGA

CGCCGAAGAAGAUGAGGACGACGCUGAGGAAGAUGAGGACGACGCCGAGGAAGAU

GACGACGACGCUGAAGAAGAUGACGACGACGCCGAAGAAGAUGAUGAUGAGGACGA

GGACGAAGAUGAGGAAGAAGAAGAAGACGAGGAAGAAGAAGAAGAAUCCGAGAAGA

AGAUCAAGCGGAACCUGCGGAAGAACGCCAAGAUCUGAUGA

305 78 AUGGGCGGAGCUGCUGCUAGACUGGGAGCCGUGAUCCUGUUCGUGGUUAUCGUG

GGACUGCAUGGCGUGCGGGGCGACAAGAGCAAAGAGCACAAGUCCAAGGGCAAGAA

GGACAAGGGGAAGAAAGAUAAGGGCAAGCACAAGAAGGCCAAGAAAGAGAAAGUCA

AGAAGCACGUCGUCAAGAACGUGAUCGAGGACGAGGACAAGGACGGCGUGGAAAUC

AUCAACCUUGAGGACAAAGAGGCCUGCGAGGAACAGCACAUCACCGUGGAAAGCAG

ACCCCUGUCUCAGCCCCAGUGCAAGCUGAUCGAUGAGCCUGAGCAGCUGACCCUGA

UGGACAAGAGCAAGGUCGAGGAAAAGCAGCUGAGCAUCCAAGAACAGCUGAUCGGC ACCAUCGGCAGAGUGAACGUGGUGCCCAGACGGGACAACCACAAGAAAAAGAUGGC

CAAGAUCGAGGAAGCCGAGCUGCAAAAGCAGAAACACGUCGACAAAGAAGAAGAUA AAGAAGAAGAGGAAGAGGAAGAAGAAGAAGAAGAGGACGAGGUCGAGGAAGAUGA

GGACGACGCCGAAGAAGAUGAGGACGACGCUGAGGAAGAUGAGGACGACGCCGAG

GAAGAUGACGACGACGCUGAAGAAGAUGACGACGACGCCGAAGAAGAUGAUGAUGA

GGACGAGGACGAAGAUGAGGAAGAAGAAGAAGACGAGGAAGAAGAAGAAGAAUCCG

AGAAGAAGAUCAAGCGGAACCUGCGGAAGAACGCCAAGAUCGGAGGCUCUGGCGGU

GGACUUAUUGCCGGCGCUGUUGGAGGAUCUCUGCUGGCCGCUCUGGUCAUCUGUG

GCAUCGUGUACUGGAUGCGGCGGCACACACAGAAGGCCCCUAAGAGAAUCAGACUG

CCCCACAUCAGAUGAUGA

[0678] In some embodiments, a GARP construct described herein has a nucleic add sequence provided in Table 24. In some embodiments, a GARP construct comprises a nucleic add sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to a nudek add sequence of Table 24. [0682] The term ”5'-cap" as used herein refers to a structure found on the S'-end of an RNA, e.g., mRNA, and generally includes a guanosine nudeotide connected to an RNA, e.g., mRNA, via a 5*- to S'-triphosphate linkage (also referred to as Gppp or G(5')ppp(5')). In some embodiments, a guanosine nudeoside included in a 5* cap may be modified, for example, by methylation at one or more positions (e.^., at the 7-position) on a base (guanine), and/or by methylation at one or more positions of a ribose. In some embodiments, a guanosine nudeoside included in a 5' cap comprises a 3'0 methylation at a ribose (3'0MeG). In some embodiments, a guanosine nucleoside included in a 5' cap comprises methylation at the 7-position of guanine (mTG). In some embodiments, a guanosine nudeoside induded in a 5' cap comprises methylation at the 7-position of guanine and a 3' 0 methylation at a ribose (m7(3'OMeG)). It will be understood that the notation used In the above paragraph, e.g., "(mz⁷-³"⁰^" or "m7(3'OMeG)", applies to other structures described herein.

[0683] In some embodiments, providing an RNA with a 5’-cap disclosed herein may be achieved by in vitro transcription, in which a S’-cap is co-transcriptlonally expressed into an RNA strand, or may be attached to an RNA post-transcriptlonally using capping enzymes. In some embodiments, co-transcriptional capping with a cap disclosed Improves the capping effidency of an RNA compared to co-transcriptional capping with an appropriate reference comparator. In some embodiments, improving capping efficiency can increase a translation effidency and/or translation rate of an RNA, and/or increase expression of an encoded polypeptide. In some embodiments, alterations to polynucleotides generates a non-hydrolyzable cap structure which can, for example, prevent decapping and increase RNA half-life.

[0684] In some embodiments, a utilized 5' caps is a capO, a capl, or cap2 structure. See, e.g., Fig. 1 of Ramanathan A et a!., and Fig. 1 of Decroly E etal, each of which is Incorporated herein by reference in Its entirety. See, e.g., Rg. 1 of Ramanathan A et al., and Rg. 1 of Decroly E eta!., each of which is incorporated herein by reference In its entirety. In some embodiments, an RNA described herein comprises a capl structure. In some embodiments, an RNA described herein comprises a cap2.

[0685] In some embodiments, an RNA described herein comprises a capO structure. In some embodiments, a capO structure comprises a guanosine nucleoside methylated at the 7-position of guanine ((m⁷^). In some embodiments, such a capO structure Is connected to an RNA via a 5'- to S'-triphosphate linkage and Is also referred to herein as (m⁷)Gppp. In some embodiments, a capO structure comprises a guanosine nucleoside methylated at the 2'-position of the ribose of guanosine. In some embodiments, a capO structure comprises a guanosine nucleoside methylated at the 3'-position of the ribose of guanosine . In some embodiments, a guanosine nucleoside included in a 5' cap comprises methylation at the 7-position of guanine and at the 2'-position of the ribose ((m₂ ⁷,^3'-0)G). In some embodiments, a guanosine nucleoside included in a 5' cap comprises methylation at the 7-position of guanine and at the 2'-position of the ribose ((m₂ ⁷,^3'-0)G).

[0686] In some embodiments, a capl structure comprises a guanosine nucleoside methylated at the 7-position of guanine ((m^G) and optionally methylated at the 2' or 3' position of the ribose, and a 2'0 methylated first nucleotide in an RNA ((m^N_i). In some embodiments, a capl structure comprises a guanosine nucleoside methylated at the 7-position of guanine ((m⁷)G) and the 3' position of the ribose, and a 2*0 methylated first nudeotide in an RNA ((m^2'-0)N1). In some embodiments, a capl structure is connected to an RNA via a 5'- to S’- triphosphate Inkage and is also referred to herein as, e.g., ((m⁷)Gppp(^2'-0)N₁) or (m₂ ^7,3'-0)Gpp(²'‘⁰)N₁), wherein N₁ is as defined and described herein. In some embodiments, a capl structure comprises a second nudeotide, N₂, which is at position 2 and is chosen from A, G, C, or U, e.g., (m⁷)Gppp(²'"⁰)N₁pN2 or (m₂ ⁷'^3'-0)Gppp(^z"⁰)N₁pN2 , wherein each of N₁ and N₂ is as defined and described herein.

[0687] In some embodiments, a cap2 structure comprises a guanosine nudeoside methylated at the 7 -position of guanine ((m⁷)G) and optionally methylated at the 2' or 3' position of the ribose, and a 2'0 methylated first and second nudeotides in an RNA ((m²'"°)N₁p(m²'"^<l)N2). In some embodiments, a cap2 structure comprises a guanosine nudeoside methylated at the 7-position of guanine ((m^G) and the 3' position of the ribose, and a 2'0 methylated first and second nucleotide in an RNA. In some embodiments, a cap2 structure is connected to an RNA via a 5'- to 5'- triphosphate inkage and is also referred to herein as, e.g., ((m⁷)Gppp(^2'-0)N₁p(^z"^o)N2) or (m2⁷-³’"⁰)Gppp(²'"°)N₁p(²'" ⁰)N₂), wherein each of N₁ and N2 is as defined and described herein.

[0688] In some embodiments, the 5' cap Is a dinucleotide cap structure. In some embodiments, the 5' cap Is a dinucleotide cap structure comprising N₁, wherein N₁ is as defined and described herein. In some embodiments, the 5' cap is a dinucleotide cap G*N₁, wherein N₁ is as defined above and herein, and G* comprises a structure of formula (I): or a salt thereof, wherein each R² and R³ is -OH or -OCH3; and X is 0 or S.

[0689] In some embodiments, R² is -OH. In some embodiments, R² is -OCH3. In some embodiments, R³ is -

OH. In some embodiments, R³ is -OCH3. In some embodiments, R² is -OH and R³ is -OH. In some embodiments, R² is -OH and R³ is -CH3. In some embodiments, R² is -CH3 and R³ is -OH. In some embodiments, R² is -CH3 and R³ is - CH₃.

[0690] In some embodiments, X is 0. In some embodiments, X is S.

[0691] In some embodiments, the 5' cap is a dinucleotide capO structure (e.g., (m7) GpppN₁, (m₂ ⁷'^2'-0)GpppN₁, (m₂ ⁷'^3'-0)GpppN₁, (m⁷)GppSpN₁, (m₂ ⁷'^2'-0)GppSpN₁, or (m₂ ⁷'^3'-0)GppSpN₁), wherein N₁ is as defined and described herein. In some embodiments, the 5' cap is a dinudeotide capo structure (e.#,, (m⁷)GpppN₁, (m₂ ⁷'^2'-0)GpppN₁, (m2⁷'³*" °)GpppN₁, (m⁷)GppSpN₁, (m₂ ⁷'^2'-0)GppSpN₁, or (m₂ ⁷'^3'-0)GppSpN₁), wherein N₁ is G. In some embodiments, the 5' cap is a dinudeotide capO structure (m^pppN₁, (m₂ ⁷'^2'-0)GpppN₁, (m₂ ⁷'^3'-0)GpppN₁, (m7) GppSpN₁, (m₂7,2'- °)GppSpN₁, or (m₂ ⁷'^3'-0)GppSpN₁), wherein N₁ is A, U, or C. In some embodiments, the 5' cap is a dinudeotide capl structure (e.g., (m⁷)Gppp(m²'‘⁰)N₁, (m₂ ⁷'^2'-0)Gppp(m^2'-0)N₁, (m₂ ⁷'^3'-0)Gppp(m^2'-0)N₁, (m⁷)GppSp(m^2'-0)N₁, (m2⁷'²'" °)GppSp(m^2'-0)N₁, or (m₂ ⁷'^3'-0)GppSp(m^2'-0)N₁), wherein N₁ is as defined and described herein. In some embodiments, the 5' cap is selected from the group consisting of (m⁷)GpppG ("EcapO"), (m⁷X3ppp(m^2'-0)G ("Ecapl”), (m2⁷-³'" °)GpppG ("ARCA" or "DI"), and (m₂ ⁷'^2'-0)GppSpG ("beta-S-ARCA”). In some embodiments, the 5' cap is (m⁷)GpppG ("EcapO"), having a structure: [0695] In some embodiments, the 5' cap is a trinucleotide cap structure. In some embodiments, the 5' cap is a trinucleotide cap structure comprising N₁pN₂, wherein N₁ and N₂ are as defined and described herein. In some embodiments, the 5' cap is a dinucleotide cap G*N₁pN₂, wherein N₁ and N₂ are as defined above and herein, and G* comprises a structure of formula (I): or a salt thereof, wherein R², R³, and X are as defined and described herein.

[0696] In some embodiments, the 5' cap Is a trinucleotide capO structure (ay. (m^GpppN₁pN₂, (mz⁷-²'" °)GpppN₁pN2, or (mz^^GpppN₁pN₂), wherein N₁ and N? are as defined and described herein). In some embodiments, the 5' cap Is a trinucleotide capl structure (ay., (m⁷)Gppp(m²’"⁰)N₁pN₂, (mz⁷'²'⁰)Gppp(m²'‘⁰)N₁pN₂, (mz⁷'³'^X3)Gppp(m²''⁰)N₁pN₂), wherein N₁ and N₂ are as defined and described herein. In some embodiments, the 5' cap is a trinucleotide cap2 structure (ay., (m⁷)Gppp(m^2'-0)N₁p(m^2'-0)N2, (mz⁷-²'"°)Gppp(m^2'-0)N₁p(m²'"^o)N₂, (mz⁷'³'" °)Gppp(m^2'-0)N₁p(m^2'-0)N₂), wherein N₁ and N₂ are as defined and described herein. In some embodiments, the 5' cap Is selected from the group consisting of (mz⁷'³'"°)Gppp(m^2'-0)ApG ("OeanCap AG", "CC413”), (mz^7,3'"°)Gppp(m²'" °)GpG ("OeanCap GG”), (m⁷)Gppp(m²'^<>)ApG, (m⁷)Gppp(m²'"°)GpG, (mz^7<3' °)Gppp(m2⁶'²'"⁰)ApG, and (m⁷)Gppp(m²'" °)ApU.

[0697] In some embodiments, the 5' cap is (m2⁷'³'“°)Gppp(m^2'-0)ApG ("OeanCap AG", "CC413"), having a structure: or a salt thereof.

[0698] In some embodiments, the 5' cap is (mz⁷'³'"°)Gppp(m²'"^c)GpG ("CleanCap GG"), having a structure: or a salt thereof.

[0699] In some embodiments, the 5' cap is (m²)Gppp(m^2'-0)ApG, having a structure: or a salt thereof.

[0700] In some embodiments, the 5' cap is (m⁷)Gppp(m^2'-0)GpG, having a structure:

or a salt thereof.

[0706] In some embodiments, the 5' cap is (m₂ ⁷'^3'-0)Gppp(m²'"^o)Gp(m²'"^o)GpC, having a structure: or a salt thereof.

[0707] In some embodiments, the 5' cap Is (m⁷)Gppp(m^2'-0)Ap(m²''⁰)UpA, having a structure:

or a salt thereof.

[0708] In some embodiments, the 5' cap is (m⁷)Gppp(m^2'-0)Ap(m^2'-0)GpG, having a structure: or a salt thereof.

2 Cap Proximal Sequences

[0709] In some embodiments, a 5' UTR utilized In accordance with the present disclosure comprises a cap proximal sequence, e.g., as disclosed herein. In some embodiments, a cap proximal sequence comprises a sequence adjacent to a 5' cap. In some embodiments, a cap proximal sequence comprises nucleotides in positions +1, +2, +3, +4, and/or +5 of an RNA polynucleotide.

[0710] In some embodiments, a cap structure comprises one or more polynucleotides of a cap proximal sequence. In some embodiments, a cap structure comprises an m⁷ Guanosine cap and nucleotide +1 (N₁) of an RNA polynucleotide. In some embodments, a cap structure comprises an m⁷ Guanosine cap and nucleotide +2 (N₂) of an RNA polynucleotide. In some embodiments, a cap structure comprises an m⁷ Guanosine cap and nucleotides +1 and +2 (N₁ and N₂) of an RNA polynucleotide. In some embodiments, a cap structure comprises an m⁷ Guanosine cap and nucleotides +1, +2, and +3 (N₁, N₂, and N₃) of an RNA polynucleotide.

[0711] Those skilled in the art, reading the present disclosure, will appreciate that, in some embodiments, one or more residues of a cap proximal sequence (e.g., one or more of residues +1, +2, +3, +4, and/or +5) may be induded in an RNA by virtue of having been included in a cap entity (e.g., a capl or cap2 structure, etc.); alternatively, in some embodiments, at least some of the residues in a cap proximal sequence may be enzymatically added (e.g., by a polymerase such as a T7 polymerase). For example, in certain exemplified embodiments where a m₂ ⁷'^3'-0Gppp(m₁ ^2'-0)ApG cap is utilized, +1 (i.e., N₁) and +2 (l.e. N₂) are the (m₁ ^2'-0)A and G residues of the cap, and +3, +4, and +5 are added by polymerase (e.g., T7 polymerase).

[0712] In some embodiments, the S' cap is a dinucleotide cap structure, wherein the cap proximal sequence comprises N₁ of the 5' cap, where N₁ is any nucleotide, e.g., A, C, G or U. In some embodiments, the 5' cap Is a trinucleotide cap structure (e.g^.._, the trinucleotide cap structures described above and herein), wherein the cap proximal sequence comprises N₁ and N₂ of the 5' cap, wherein N₁ and N₂ are Independently any nucleotide, e.g., A, C, G or U. In some embodiments, the 5' cap Is a tetranucleotide cap structure (e.g., the trinucleotide cap structures described above and herein), wherein the cap proximal sequence comprises N₁, N₂, and N3 of the 5' cap, wherein N₁, N₂, and N? are any nucleotide, e.g., A, C, G or U.

[0713] In some embodiments, e.g., where the 5' cap Is a dinucleotide cap structure, a cap proximal sequence comprises N₁ of a the 5' cap, and N₂, N3, N₁ and Ns, wherein N₁ to Ns correspond to positions +1, +2, +3, +4, and/or +5 of an RNA polynucleotide. In some embodiments, e.g., where the 5' cap is a trinucleotide cap structure, a cap proximal sequence comprises N₁ and N₂ of a the 5' cap, and N₃, N₄ and N₅, wherein N₁ to N₅ correspond to positions +1, +2, +3, +4, and/or +5 of an RNA polynucleotide. In some embodiments, e.g., where the 5' cap is a tetranudeotide cap structure, a cap proximal sequence comprises N₁, N₂, and N₃ of a the 5' cap, and N₁ and Ns, wherein N₁ to Ns correspond to positions +1, +2, +3, +4, and/or +5 of an RNA polynucleotide.

[0714] In some embodiments, N₁ is A. In some embodiments, N₁ is C. In some embodiments, N₁ is G. In some embodiments, N₁ is U. In some embodiments, N₂ is A In some embodiments, N₂ is C. In some embodiments, N₂ is G. In some embodiments, N₂ is U. In some embodiments, N3 Is A In some embodiments, N3 is C. In some embodiments, N3 is G. In some embodiments, N₃ is U. In some embodiments, N₁ is A. In some embodiments, N₁ is C. In some embodiments, N₁ is G. In some embodiments, N₁ is U. In some embodiments, Ns is A. In some embodiments, Ns is C. In some embodiments, Ns is G. In some embodiments, N₅ is U. It will be understood that, each of the embodiments described above and herein (e.g., for N₁ through Ns) may be taken singly or in combination and/or may be combined with other embodiments of variables deserved above and herein (e.g., 5' caps).

[0715] In some embodiments, a cap proximal sequence comprises Ai and G₂ of the Capl structure, and a sequence comprising: A₃A₄U₅ (SEQ ID NO: 138) at positions +3, +4 and +5 respectively of the polyribonucleotide.

3. 5'UTR [0716] In some embodiments, a nucleic add (e.g., DNA, RNA) utilized in accordance with the present disclosure comprises a 5* -UTR. In some embodiments, 5'-UTR may comprise a plurality of distinct sequence elements; in some embodiments, such plurality may be or comprise multiple copies of one or more particular sequence elements (e.g., as may be from a particular source or otherwise known as a functional or characteristic sequence element). In some embodiments a 5' UTR comprises multiple different sequence elements.

[0717] The term "untranslated region" or "UTR" is commonly used in the art to a region in a DNA molecule which is transcribed but is not translated into an amino add sequence, or to the corresponding region in an RNA pdynudeotide, such as an mRNA molecule. An intranslated region (UTR) can be present 5' (upstream) of an open reading frame (5'-UTR) and/or 3' (downstream) of an open reading frame (3'-UTR). As used herein, the terms "five prime untranslated region" or "S' UTR" refer to a sequence of a polyribonudeotide between the 5' end of the polyribonucleotide (e.g., a transcription start site) and a start codon of a coding region of the polyribonucleotide. In some embodiments, "5' UTR” refers to a sequence of a polyribonudeotide that begins at the 5' end of the polyribonudeotide (e.g., a transcription start site) and ends one nucleotide (nt) before a start codon (usually AUG) of a coding region of the polyribonucleotide, e.g., in Its natural context. In some embodiments, a 5' UTR comprises a Kozak sequence. A 5*-UTR Is downstream of the S'-cap (if present), e.g., directly adjacent to the S'-cap. In some embodiments, a 5' UTR disdosed herein comprises a cap proximal sequence, e.g., as defined and described herein. In some embodiments, a cap proximal sequence comprises a sequence adjacent to a 5' cap.

[0718] Exemplary 5' UTRs indude a human alpha globin (hAg) S'UTR or a fragment thereof, a TEV 5' UTR or a fragment thereof, a HSP70 5' UTR or a fragment thereof, or a c-Jun 5' UTR or a fragment thereof.

[0719] In some embodiments, an RNA disclosed herein comprises a hAg 5' UTR or a fragment thereof. [0720] In some embodiments, an RNA disclosed herein comprises a 5' UTR having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% Identity to a 5' UTR with the sequence AGAATAAACTAGTATTCTTCTGGTCCCCACAGACTCAGAGAGAACCCGCCACC (SEQ ID NO: 110). In some embodiments, an RNA disclosed herein comprises a 5' UTR having the sequence AGAATAAACTAGTATTCTTCTGGTCCCCACAGACTCAGAGAGAACCCGCCACC (SEQ ID NO: 110).

[0721] In some embodiments, an RNA disclosed herein comprises a 5' ITTR having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% Identity to a 5' UTR with the sequence AACUAGUAUUCUUCUGGUCCCCACAGACUCAGAGAGAACCCGCCACC (SEQ ID NO: Hl)(hAg-Kozak/5'UTR). In some embodiments, an RNA disclosed herein comprises a 5' UTR having the sequence AACUAGUAUUCUUCUGGUCCCCACAGACUCAGAGAGAACCCGCCACC (SEQ ID NO: Ul)(hAg-Kozak/5'UTR).

4. PolyA TaS

[0722] In some embodiments, a polynucleotide (e.g., DNA, RNA) disclosed herein comprises a polyadenylate (polyA) sequence, e.g., as described herein. In some embodiments, a polyA sequence is situated downstream of a 3'- UTR, e.g., adjacent to a 3' UTR.

[0723] As used herein, the term "poly(A) sequence" or "poly-A tail" refers to an uninterrupted or interrupted sequence of adenylate residues which is typically located at the 3'-end of an RNA polynucleotide. Poly(A) sequences are known to those of skill in the art and may follow the 3'-UTR in the RNAs described herein. An uninterrupted poly(A) sequence is characterized by consecutive adenylate residues. In nature, an uninterrupted poly(A) sequence is typical. In some embodiments, polynucleotides disclosed herein comprise an uninterrupted Poly(A) sequence. In some embodiments, polynucleotides disclosed herein comprise interrupted Poly(A) sequence. In some embodiments, RNAs disclosed herein can have a poly(A) sequence attached to the free 3'-end of the RNA by a templateindependent RNA polymerase after transcription or a poly(A) sequence encoded by DNA and transcribed by a template-dependent RNA polymerase.

[0724] It has been demonstrated that a pdy(A) sequence of about 120 A nucleotides has a beneficial influence on the levels of RNA in transfected eukaryotic cells, as well as on the levels of protein that is translated from an open reading frame that is present upstream (5*) of the poly(A) sequence (Holtkamp et al, 2006, Blood, vol. 108, pp. 4009-4017, which is herein incorporated by reference).

[0725] In some embodiments, a poly(A) sequence in accordance with the present disclosure is not limited to a particular length; in some embodiments, a poly(A) sequence is any length. In some embodiments, a poly(A) sequence comprises, essentially consists of, or consists of at least 20, at least 30, at least 40, at least 80, or at least 100 and up to 500, up to 400, up to 300, up to 200, or up to 150 A nucleotides, and, in particular, about 120 A nucleotides. In this context, "essentially consists of" means that most nucleotides in the poly(A) sequence, typically at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% by number of nucleotides in the poiy(A) sequence are A nucleotides, but permits that remaining nucleotides are nucleotides other than A nucleotides, such as U nucleotides (uridylate), G nucleotides (guanylate), or C nucleotides (cytidylate). In this context, "consists of" means that all nucleotides in the poly(A) sequence, i.e., 100% by number of nucleotides in the poly(A) sequence, are A nucleotides. The term "A nucleotide" or "A" refers to adenylate.

[0726] In some embodiments, a poly(A) sequence is attached during RNA transcription, e.g., during preparation of in vitro transcribed RNA, based on a DNA template comprising repeated dT nucleotides (deoxythymidylate) in the strand complementary to the coding strand. The DNA sequence encoding a poly(A) sequence (coding strand) Is referred to as poly(A) cassette.

[0727] In some embodiments, the poly(A) cassette present in the coding strand of DNA essentially consists of dA nucleotides, but Is interrupted by a random sequence of the four nucleotides (dA, dC, dG, and dT). Such random sequence may be 5 to 50, 10 to 30, or 10 to 20 nucleotides In length. Such a cassette is disclosed in WO 2016/005324 Al, hereby Incorporated by reference. Any poly(A) cassette disclosed in WO 2016/005324 Al, which is incorporated herein by reference in Its entirety, may be used In accordance with the present disclosure. A poly(A) cassette that essentially consists of dA nucleotides, but is interrupted by a random sequence having an equal distribution of the four nucleotides (dA, dC, dG, dT) and having a length of e.g., 5 to 50 nucleotides shows, on DNA level, constant propagation of plasmid DNA in £ atf/'and is still associated, on RNA level, with the beneficial properties with respect to supporting RNA stability and translational efficiency is encompassed. In some embodiments, the poly(A) sequence contained in an RNA polynucleotide described herein essentially consists of A nucleotides, but is interrupted by a random sequence of the four nucleotides (A, C, G, U). Such random sequence may be 5 to 50, 10 to 30, or 10 to 20 nucleotides in length.

[0728] In some embodiments, no nucleotides other than A nucleotides flank a poly(A) sequence at its 3'-end, i.e., the poly(A) sequence is not masked or followed at its 3'-end by a nucleotide other than A.

[0729] In some embodiments, the poly(A) sequence may comprise at least 20, at least 30, at least 40, at least 80, or at least 100 and up to 500, up to 400, up to 300, up to 200, or up to 150 nucleotides. In some embodiments, the poly(A) sequence may essentially consist of at least 20, at least 30, at least 40, at least 80, or at least 100 and up to 500, up to 400, up to 300, up to 200, or up to 150 nucleotides. In some embodiments, the poly(A) sequence may consist of at least 20, at least 30, at least 40, at least 80, or at least 100 and up to 500, up to 400, up to 300, up to 200, or up to 150 nucleotides. In some embodiments, the poly(A) sequence comprises at least 100 nucleotides. In some embodiments, the poly(A) sequence comprises about 150 nucleotides. In some embodiments, the poly(A) sequence comprises about 120 nucleotides.

[0730] In some embodiments, a poly A tail comprises a specific number of Adenosines, such as about 50 or more, about 60 or more, about 70 or more, about 80 or more, about 90 or more, about 100 or more, about 120, or about 150 or about 200. In some embodiments a poly A tail of a string construct may comprise 200 A residues or less. In some embodiments, a poly A tail of a string construct may comprise about 200 A residues. In some embodiments, a poly A tall of a string construct may comprise 180 A residues or less. In some embodiments, a poly A tail of a string construct may comprise about 180 A residues. In some embodiments, a poly A tall may comprise 150 residues or less.

[0731] In some embodiments, RNA comprises a poly(A) sequence comprising the nucleotide sequence of AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAGCATATGACTAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

AAAAAAAAAAAAAAAAAAAAAAAA (SEQ ID NO: 112), or a nucleotide sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% Identity to the nucleotide sequence of

AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAGCATATGACTAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

AAAAAAAAAAAAAAAAAAAAAAAA (SEQ ID NO: 112). In some embodiments, a poly(A) tai! comprises a plurality of A residues interrupted by a linker. In some embodiments, a linker comprises the nucleotide sequence GCATATGAC (SEQ ID NO: 113).

[0732] In some embodiments, RNA comprises a poly(A) sequence comprising the nucleotide sequence of AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAGCAUAUGACUAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

AAAAAAAAAAAAAAAAAAAAAAAA (SEQ ID NO: 114), or a nucleotide sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% Identity to the nucleotide sequence of

AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAGCAUAUGACUAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

AAAAAAAAAAAAAAAAAAAAAAAA (SEQ ID NO: 114).

[0733] In some embodiments, a poly(A) tail comprises a plurality of A residues interrupted by a linker. In some embodiments, a linker comprises the nucleotide sequence GCAUAUGAC (SEQ ID NO: 115).

5. 3' UTR

CTGCCACCTCCACCTGCCCCACTCACCACCTCTGCTAGTTCCAGACACCTCCCAAGCACGCAGCAATGCAGCTCAAAACGCTTAG

CCTAGCCACACCCCCACGGGAAACAGCAGrGATTAACCTTTAGCAATAAACGAAAGTTTAACTAAGCTATACTAACCCCAGGGTT

GGTCAATTTGGTGCCAGCCACACC (SEQ ID NO: 116). In some embodiments, an RNA disclosed herein comprises a 3' UTR with the sequence of

CTGGTACTGCATGCACGCAATGCTAGCTGCCCCTTTCCCGTCCTGGGTACCCCGAGTCTCCCCCGACCTCGGGrCCCAGGTATG

CTCCCACCTCCACCTGCCCCACTCACCACCTCTGCTAGTTCCAGACACCTCCCAAGCACGCAGCAATGCAGCTCAAAACGCTTAG

CCTAGCCACACCCCCACGGGAAACAGCAGTGATTAACCTTTAGCAATAAACGAAAGTTTAACTAAGCTATACTAACCCCAGGGTT

GGTCAATTTCGTGCCAGCCACACC (SEQ ID NO: 116).

[0738] In some embodiments, an RNA disclosed herein comprises a 3' UTR having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to a 3' UTR with the sequence of

CUGGUACUGCAUGCACGCAAUGCUAGCUGCCCCUUUCCCGUCCUGGGUACCCCGAGJCUCCCCCGACCUCGGGUCCCAGGUA

UGCUCCCACCUCCACCUGCCCCACUCACCACCUCUGCUAGUUCCAGACACCUCCCAAGCACGCAGCAAUGCAGCUCAAAACGCU

UAGCCUAGCCACACCCCCACGGGAAACAGCAGUGAUUAACCUUUAGCAAUAAACGAAAGUUUAACUAAGCUAUACUAACCCCA

GGGUUGGUCAAUUUCGUGCCAGCCACACC (SEQ ID NO: 117).

[0739] In some embodiments, an RNA disclosed herein comprises a 3' UTR with the sequence of CUGGUACUGCAUGCACGCAAUGCUAGCUGCCCCUUUCCCGUCCUGGGUACCCCGAGUCUCCCCCGACCUCGGGUCCCAGGUA

GGGUUGGUCAAUUUCGUGCCAGCCACACC (SEQ ID NO: 117).

[0741] At least three distinct formats useful for RNA compositions (e.g., pharmaceutical compositions) have been developed, namely non-modified uridine containing RNA (uRNA), nucleoside-modified RNA (modRNA), and selfamplifying RNA (saRNA). Each of these platforms displays unique features. In general, in all three formats, RNA Is capped, contains open reading frames (ORFs) flanked by untranslated regions (UTR), and have a polyA-tail at the 3' end. An ORF of an uRNA and modRNA vectors encode an antibody agent or portion thereof. An saRNA has multiple ORFs.

[0742] In some embodiments, the RNA described herein may have modified nucleosides. In some embodiments, the RNA comprises a modified nucleoside in place of at least one (e.g., every) uridine.

[0743] The term "uracil," as used herein, describes one of the nudeobases that can occur in the nucleic acid of RNA. The structure of uracil is: o

NH

N ■o H

[0744] The term "uridine," as used herein, describes one of the nucleosides that can occur in RNA. The structure of uridine Is:

[0745] UTP (uridine 5'-triphosphate) has the following structure:

[0746] Pseudo-UTP (pseudouridine 5'-triphosphate) has the following structure: O

HN NH

O

.0.

OH OH

[0747] "Pseudouridine" is one example of a modified nucleoside that is an isomer of uridine, where the uracil is attached to the pentose ring via a carbon-carbon bond instead of a nitrogen-carbon glycosidic bond.

[0748] Another exemplary modified nucleoside is Nl-methyl-pseudouridine (mlU>), which has the structure:

[0749] Nl-methyl-pseudo-UTP has the following structure:

[0750] Another exemplary modified nucleoside is 5-methyl-uridlne (m5U), which has the structure:

[0751] In some embodiments, one or more uridine in the RNA described herein is replaced by a modified nucleoside. In some embodiments, the modified nucleoside is a modified uridine.

[0752] In some embodiments, RNA comprises a modified nucleoside in place of at least one uridine. In some embodiments, RNA comprises a modified nucleoside in place of each uridine.

[0753] In some embodiments, the modified nucleoside is independently selected from pseudouridine (Ψ ), Nl- methyl-pseudoiridine (mlΨ), and 5-methyl-uridine (m5U). In some embodiments, the modified nucleoside comprises pseudouridine (Ψ). In some embodiments, the modified nucleoside comprises Nl-methyl-pseudouridine (mlΨ). In some embodiments, the modified nucleoside comprises 5-methyl-uridine (m5U). In some embodiments, RNA may comprise more than one type of modified nucleoside, and the modified nucleosides are independently selected from pseudouridine (Ψ), Nl-methyl-pseudouridine (m1Ψ ), and 5-methyl-uridine (m5U). In some embodiments, the modified nucleosides comprise pseudouridine (Ψ ) and Nl-methyl-pseudouridine (mlΨ ). In some embodiments, the modified nucleosides comprise pseudouridine (Ψ ) and 5-methyl-uridlne (m5U). In some embodiments, the modified nucleosides comprise Nl-methyl-pseudouridine (mlΨ ) and 5-methyl-uridlne (m5U). In some embodiments, the modified nucleosides comprise pseudouridine (Ψ ), Nl-methyl-pseudouridine (mlΨ ), and 5-methyl-uridine (m5U). [0754] In some embodiments, the modified nudeoside replacing one or more, e.g., all, uridine in the RNA may be any one or more of 3-methyl-uridine (m3U), 5-methoxy-uridine (mo5U), 5-aza-uridine, 6-aza-uridine, 2-thio-5- aza-uridine, 2-thio-uridine (s2U), 4-thio-uridine (s4U), 4-thio-pseudouridine, 2-thio-pseudouridine, 5-hydroxy-uridine (ho5U), 5-aminoafiyl-uridine, 5-halo-uridine (e.g., 5-iodo-uridine or 5 -bromo-uridine), uridine 5-oxyacetic acid (cmo5U), uridine 5-oxyacetic add methyl ester (mcmo5U), 5-carboxymethyl-uridine (cm5U), 1 -carboxymethylpseudouridine, 5-carboxyhydroxymethyl-uridine (chm5U), 5-carboxyhydroxymethyl-uridine methyl ester (mchm5U), 5-methoxycarbonylmethyl-uridine (mcm5U), 5-methoxycarbonylmethyl-2-thio-uridine (mcm5s2U), 5-aminomethyl-2- thio-uridine (nm5s2U), 5-methylaminomethyl-uridine (mnm5U), 1-ethyl-pseudouridine, 5-methylaminomethyl-2-thio- uridine (mnm5s2U), 5-methylamlnomethyl-2-seleno-uridlne (mnm5se2U), 5-carbamoylmethyl-urldine (ncm5U), 5- carboxymethylaminomethyl-urldlne (cmnm5U), 5-carboxymethylaminomethyl-2-thio-uridine (cmnm5s2U), 5- propynyl-uridine, 1-propynyl-pseudouridlne, 5-taurinomethyl-urldine (τm5U ), 1-taurinomethyl-pseudouridine, 5- taurinomethyl-2-thk>-uridine(τm 5s2U), l-taurinomethyl-4-thio-pseudouridine), 5-methyl-2-thio-uridine (m5s2U), 1- methyl-4-thlo-pseudouridine (mls4Ψ ), 4-thio-l-methyl-pseudouridine, 3-methyl-pseudouridine (m3Ψ), 2-thio-l- methyl-pseudouridine, 1-methyl-l-deaza-pseudouridlne, 2-thio-l-methyl-l-deaza-pseudouridlne, dihydrouridine (D), dihydropseudouridine, 5,6-dihydrouridine, 5-methyl-dihydrouridine (m5D), 2-thio-dihydrouridlne, 2-thio- dihydropseudouridine, 2-methoxy-uridine, 2-methoxy-4-thio-uridine, 4-methoxy-pseudouridlne, 4-methoxy-2-thio- pseudouridine, Nl-methyl-pseudouridine, 3-(3-amlno-3-carboxypropyl)uridlne (acp3U), l-methyl-3-(3-amlno-3- carboxypropyl)pseudouridlne (acp3 Ψ ), 5-(isopentenylamlnomethyl)uridine (in 5U), 5-(isopentenylaminomethyl)-2- thlo-uridlne (inm5s2U), α-thio-uridlne, 2'-0-methyl-uridlne (Um), 5,2'-O-dimethyl-uridine (m5Um), 2'-0-methyl- pseudouridine (Ψ m), 2-thio-2'-O-methyl-uridine (s2Um), 5-methoxycarbonylmethyl-2'-O-methyl-urldine (mcm5Um), 5-carbamoylmethyl-2'-O-methyl-uridine (ncm5Um), 5-carboxymethylamlnomethyl-2'-O-methyl-uridlne (cmnm5Um), 3,2'-O-dimethyl-uridine (m3Um), 5-(lsopentenylaminomethyl)-2'-O-methyl-uridine (Inm5Um), 1-thio-urldlne, deoxythymidine, 2'-F-ara-ur1dlne, 2'-Furidine, 2'-0H-ara-uridine, 5-(2-carbomethoxyvlnyl) uridine, 5-[3-(l-E- propenylamino)uridine, or any other modified uridine known In the art [0755] In some embodiments, the RNA comprises other modified nucleosides or comprises further modified nucleosides, e.g., modified cytidine. For example, In some embodiments, in the RNA 5-methylcytidine is substituted partially or completely, preferably completely, for cytidine. In some embodiments, the RNA comprises 5- methykytidine and one or more selected from pseudouridne (Ψ ), Nl-methyl-pseudouridine (mlΨ), and 5-methyl- uridine (m5U). In some embodiments, the RNA comprises 5-methylcytidine and Nl-methyl-pseudouridine (mlΨ). In some embodiments, the RNA comprises 5-methylcytidine in place of each cytidine and Nl-methyl-pseudouridine (mlΨ) in place of each uridine.

[0756] In some embodiments of the present disclosure, the RNA is "replicon RNA" or simply a "replicon," in particular "self-replicating RNA" or "self-amplifying RNA.” In one particularly preferred embodiment, the replicon or self-replicating RNA is derived from or comprises elements derived from a single-stranded (ss) RNA virus, in particular a positive-stranded ssRNA virus, such as an alphavirus. Alphaviruses are typical representatives of positive- stranded RNA viruses. Alphaviruses replicate in the cytoplasm of infected cells (for review of the alphaviral life cycle see José et al., Future Microbiol., 2009, vol. 4, pp. 837-856, which is incorporated herein by reference in its entirety). The total genome length of many alphaviruses typically ranges between 11,000 and 12,000 nucleotides, and the genomic RNA typically has a 5'-cap, and a 3' poly(A) tail. The genome of alphaviruses encodes non-structural proteins (involved in transcription, modification and replication of viral RNA and in protein modification) and structural proteins (forming the virus partide). There are typically two open reading frames (ORFs) in the genome. The four non-structural proteins (nsPl-nsP4) are typically encoded together by a first ORF beginning near the 5' terminus of the genome, while alphavirus structural proteins are encoded together by a second ORF which is found downstream of the first ORF and extends near the 3' terminus of the genome. Typically, the first ORF is larger than the second ORF, the ratio being roughly 2:1. In cells infected by an alphavirus, only the nucleic add sequence encoding non-structural proteins is translated from the genomic RNA, while the genetic information encoding structural proteins is translatable from a subgenomic transcript, which is an RNA molecule that resembles eukaryotic messenger RNA (mRNA; Gould et al., 2010, Antiviral Res., vol. 87 pp. 111-124, which is incorporated herein by reference In Its entirety). Following infection, i.e. at early stages of the viral life cyde, the (+) stranded genomic RNA directly acts like a messenger RNA for the translation of the open reading frame encoding the non-structural polyprotein (nsP1234).

[0757] Alphavirus-derived vectors have been proposed for delivery of foreign genetic information into target cels or target organisms. In simple approaches, a first ORF encodes an alphavirus-derived RNA-dependent RNA polymerase (replicase), which upon translation mediates self-amplification of the RNA. A second ORF encoding alphaviral structural proteins is replaced by an open reading frame encoding a Plasmodium polypeptide construct described herein. Alphavirus-based trans-replication systems rely on alphavirus nucleotide sequence elements on two separate nudeic add molecules: one nucleic acid molecule encodes a viral replicase, and the other nucleic add molecule is capable of being replicated by said replicase in trans (hence the designation trans-replication system). Trans-replication requires the presence of both these nucleic acid molecules In a given host cell. The nucleic add molecule capable of being replicated by the replicase In trans must comprise certain alphaviral sequence elements to allow recognition and RNA synthesis by the alphaviral replcase.

[0758] Features of a non-modified uridine platform may include, for example, one or more of Intrinsic adjuvant effect, as well as good tolerability and safety. Features of modified uridine (e.g., pseudouridine) platform may Include reduced adjuvant effect, blunted Immune Innate Immune sensor activating capadty and thus good tolerability and safety. Features of self-amplifying platform may indude, for example, long duration of protein expression, good tolerability and safety, higher likelihood for efficacy with very low vaccine dose.

[0759] The present disdosure provides particular RNA constructs optimized, for example, for improved manufacturability, encapsulation, expression level (and/or timing), etc. Certain components are discussed below, and certain preferred embodiments are exemplified herein.

C. Codon Optimization and GC Enrichment

[0760] As used herein, the term 'codon-optimized" refers to alteration of codons in a coding region of a nucleic add molecule (e.g., a polyribonucleotide) to reflect the typical codon usage of a host organism (e.g., a subject receiving a nudeic add molecule (e.g., a polyribonudeotkle)) without preferably altering the amino add sequence encoded by the nudeic add molecule. Within the context of the present disdosure, in some embodiments, coring regions are codon-optimized for optimal expression in a subject to be treated using the RNA molecules described herein. In some embodiments, codon-optimization may be performed such that codons for which frequently occurring tRNAs are available are inserted in place of 'rare codons." In some embodiments, codon-optimization may indude increasing guanosine/cytosine (G/C) content of a coding region of RNA described herein as compared to the G/C content of the corresponding coding sequence of a wild type RNA, wherein the amino add sequence encoded by the RNA is preferably not modified compared to the amino acid sequence. [0761] In some embodiments, a coding sequence (also referred to as a "coding region'*) is codon optimized for expression in the subject to whom a composition (e.g., a pharmaceutical composition) is to be administered (e.#,, a human). Thus, in some embodiments, sequences in such a polynucleotide (e.g._z a polyribonucleotide) may differ from wild type sequences encoding the relevant antigen or fragment or epitope thereof, even when the amino add sequence of the antigen or fragment or epitope thereof is wild type.

[0762] In some embodiments, strategies for codon optimization for expression in a relevant subject (e.^._z a human), and even, in some cases, for expression in a particular cell or tissue.

[0763] Various species exhibit particular bias for certain codons of a particular amino add. Without wishing to be bound by any one theory, codon bias (differences in codon usage between organisms) often correlates with the efficiency of translation of messenger RNA (mRNA), which Is in turn believed to be dependent on, among other things, the properties of the codons being translated and the availability of particular transfer RNA (tRNA) molecules. The predominance of selected tRNAs in a ceH may generally be a reflection of the codons used most frequently in peptide synthesis. Accordingly, genes may be tailored for optimal gene expression in a given organism based on codon optimization. Codon usage tables are available, for example, at the "Codon Usage Database" avaiable at www.kazusa.orjp/codon/ and these tables may be adapted in a number of ways. Computer algorithms for codon optimizing a particular sequence for expression in a particular subject or its cells are also available, such as Gene Forge (Aptagen; Jacobus, PA), are also available.

[0764] In some embodiments, a polynucleotide (e.y_v a polyribonucleotide) of the present disclosure is codon optimized, wherein the codons in the polynucleotide (e.^., the polyribonucleotide) are adapted to human codon usage (herein referred to as "human codon optimized polynucleotide"). Codons encoding the same amino add occur at different frequencies in a subject, e.g., a human. Accordingly, in some embodiments, the coding sequence of a polynucleotide of the present disclosure Is modified such that the frequency of the codons encoding the same amino add corresponds to the naturally occurring frequency of that codon according to the human codon usage, e.g., as shown in Table 25. For example, in the case of the amino add Ala, the wild type coding sequence Is preferably adapted in a way that the codon "GCC" is used with a frequency of 0.40, the codon "GCT" is used with a frequency of 0.28, the codon "GCA" is used with a frequency of 0.22 and the codon "GCG” Is used with 30 a frequency of 0.10 etc. (see Table 25). Accordingly, In some embodiments, such a procedure (as exemplified for Ala) is applied for each amino acid encoded by the coding sequence of a polynucleotide to obtain sequences adapted to human codon usage.

Table 25: Human codon usage table with frequencies indicated for each amino add.

Amino add codon frequency Amino add codon frequency

Ala GCG 0.10 Pro CCG 0.11

Ala GCA 0.22 Pro CCA 0.27

Ala GCT 0.28 Pro CCT 0.29

Ala GCC* 0.40 Pro CCC* 0.33

Cys TGT 0.42 Gin GAG* 0.73

Cys TGC* 0.58 Gin CAA 0.27

Asp GAT 0.44 Arg AGG 0.22

Asp GAC* 0.56 Arg AGA* 0.21

[0765] Certain strategies for codon optimization and/or G/C enrichment for human expression are described in W02002/098443, which is incorporated by reference herein in its entirety. In some embodiments, a coding sequence may be optimized using a multiparametric optimization strategy. In some embodiments, optimization parameters may include parameters that influence protein expression, which can be, for example, impacted on a transcription level, an mRNA level, and/or a translational level. In some embodiments, exemplary optimization parameters include, but are not limited to transcription-level parameters (including, e.g., GC content; consensus splice sites, cryptic splice sites, SD sequences, TATA boxes, termination signals, artificial recombination sites, and combinations thereof); mRNA-level parameters (Including, e.g., RNA instability motifs, ribosomal entry sites, repetitive sequences, and combinations thereof); translation-level parameters (Including, e.g., codon usage, premature poly(A) sites, ribosomal entry sites, secondary structures, and combinations thereof); or combinations thereof. In some embodiments, a coding sequence may be optimized by a GeneOptimizer algorithm as described in Fath et al. "Multiparameter RNA and Codon Optimization: A Standardized Tool to Assess and Enhance Autologous Mammalian Gene Expression' PLoS ONE 6(3): el7596; Rabb et al., which Is Incorporated herein by reference In Its entirety, "The GeneOptimizer Algorithm: using a sliding window approach to cope with the vast sequence space in multiparameter DMA sequence optimization" Systems and Synthetic Biology (2010) 4:215-225; and Graft et al. "Codon-optimized genes that enable increased heterologous expression in mammalian cells and elicit efficient immune responses in mice after vaccination of naked DMA" Methods Mol Med (2004) 94:197-210, the entire content of each of which is incorporated herein for the purposes described herein. In some embodiments, a coding sequence may be optimized by Eurofins' adaption and optimization algorithm "GENEius" as described in Eurofins' Application Notes: Eurofins' adaption and optimization software "GENEius" in comparison to other optimization algorithms, the entire content of which is incorporated by reference for the purposes described herein.

[0766] In some embodiments, a coding sequence utilized in accordance with the present disclosure has G/C content that is increased compared to a wild type coding sequence for a

Plasmodium construct described herein, or a portion thereof. In some embodiments, guanosine/cytidlne (G/C) content of a coding region is modified relative to a wild type coring sequence for a Plasmodium construct described herein, but the amino add sequence encoded by the polyribonucleotide not modified.

[0767] Without wishing to be bound by any particular theory, it is proposed that GC enrichment may improve translation of a payload sequence. Typically, sequences having an increased G (guanosine)/C (cytidine) content are more stable than sequences having an increased A (adenosine)/!! (uridine) content. In respect to the fact that several codons code for one and the same amino add (so-called degeneration of the genetic code), the most favorable codons for the stability can be determined (so-called alternative codon usage). Depending on the amino add to be encoded by a polyribonucleotide, there are various possibilities for modification of the ribonucleic add sequence, compared to Its wHd type sequence. In particular, codons which contain A and/or U nucleosides can be modified by substituting these codons by other codons, which code for the same amino adds but contain no A and/or U or contain a lower content of A and/or U nucleosides.

[0768] In some embodiments, G/C content of a coding region of a polyribonucleotide described herein is increased by at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, or even more compared to the G/C content of the coding region prior to codon optimization, e.g., of the wiki type RNA. In some embodiments, G/C content of a coding region of a polyribonudeotide described herein is decreased by at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, or even more compared to the G/C content of the coding region prior to codon optimization, e.g., of the wild type RNA.

[0769] In some embodiments, stability and translation efficiency of a polyribonudeotide may incorporate one or more elements established to contribute to stability and/or translation efficiency of the polyribonucleotide; exemplary such elements are described, for example, in PCT/EP2006/009448 incorporated herein by reference. In some embodiments, to increase expression of a polyribonudeotide used according to the present disclosure, a polyribonucleotide may be modified within the coding region, i.e., the sequence encoding the expressed peptide or protein, without altering the sequence of the expressed peptide or protein, for example so as to increase the GC- content to increase mRNA stability and/or to perform a codon optimization and, thus, enhance translation in cells.

D. Embodiments of polyribonucleotides encoding Plasmodium polypeptide constructs [0770] In the following, exemplary embodiments of polyribonucleotides encoding Plasmodium polypeptide constructs are described, wherein certain terms used when describing elements thereof have the following meanings: [0771] cap: 5'-cap structure, e.g., selected from the group consisting of m27,2'OG(59ppSp(5')G (in particular its DI diastereomer), m27,3'OG(5')ppp(5')G, and m27,3'-OGppp(ml2'-O)ApG. [0772] hAg-Kozak: S'-LTTR sequence of the human alpha-globin mRNA with an optimized ‘Kozak sequence' to increase translational efficiency.

[0773] sec: Sequences encoding a secretory signal. [0774] Antigen: Sequences encoding one or more Plasmodium polypeptide constructs or portions or variants thereof (e.g., immunogenic fragments of the Plasmodium polypeptide constructs or the immunogenic variants thereof), from Plasmodium faidparum, preferably Plasmodium faidparum isolate 3D7.

[0775] TMD: Sequences encoding a transmembrane region. [0776] Multimerization: Sequences encoding a multimerization region. [0777] Self-assem: self-assembling nanoparticle region. [0778] Linker: Sequences coding for peptide linkers. [0779] FI element: The 3'-UTR is a combination of two sequence elements derived from the "amino terminal enhancer of split” (AES) mRNA (called F) and the mitochondrial encoded 12S ribosomal RNA (called I). These were Identified by an ex vivo selection process for sequences that confer RNA stability and augment total protein expression.

[0780] A30L70: A poly(A)-tail measuring 110 nucleotides in length, consisting of a stretch of 30 adenosine residues, followed by a 10 nucleotide linker sequence and another 70 adenosine residues designed to enhance RNA stability and translational efficiency In dendritic cells.

[0781] In some embodiments, a polyribonucleotide encoding a Plasmodium polypeptide construct described herein has one of the following structures: cap-hAg-Kozak-Antigen-FI-A30L70 cap-hAg-Kozak-sec-Antigen-FI-A30L70

- cap-hAg-Kozak-Antigen-TMD-FI-A30L70 cap-hAg-Kozak-sec-Antigen-TMD-FI-A30L70

- cap-hAg-Kozak-Antigen-Unker-TMD-FI-A30L70 cap-hAg-Kozak-sec-Antigen- Unker-TMD-FI-A30L70 cap-hAg-Kozak-Antigen-Multimerizatlon-FI-A30L70 cap-hAg-Kozak-sec-Antigen- Multimerization -FI-A30L70 cap-hAg-Kozak-Antigen-Linker-Multimerization-FI-A30L70 cap-hAg-Kozak-sec-Antigen-Linker-Multimerization -FI-A30L70 cap-hAg-Kozak-Antigen-Self-assem-FI-A30L70 cap-hAg-Kozak-sec-Antigen-Self-assem-FI-A30L70 cap-hAg-Kozak-Antigen-Unker-Self-assem-FI-A30L70 cap-hAg-Kozak-sec-Antigen-Linker-Self-assem-FI-A30L70

[0782] In some embodiments, hAg-Kozak comprises the nucleotide sequence of SEQ ID NO: 111. In some embodiments, FI comprises the nudeotide sequence of SEQ ID NO: 117. In some embodiments, A30L70 comprises the nucleotide sequence of SEQ ID NO: 114.

[0783] In some embodiments, the secretory signal is from Plasmodium faidparum, preferably Plasmodium falciparum isolate 3D7. In some embodiments, a secretory signal is from a Rh5 invasion complex polypeptide of Plasmodium faidparum (e.g., isolate 3D7). In some embodiments, a secretory signal is from a Rh5 polypeptide of Plasmodium faidparum. In some embodiments, a secretory signal is from a CSP polypeptide of Plasmodium falciparum and has the sequence as defined in Table 3 (SEQ ID NO: 41). In some embodiments, the secretory signal is heterologous and selected from the amino add sequences as defined in Table 3. In some embodiments, the secretory signal is HSVl-gD and referred to hereto as HSV-1gDsec and has the sequence as defined in Table 3 (SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 42).

[0784] In some embodiments, the TMD is a glycosylphosphatidylinositol (GPI) anchor region from Plasmodium CSP, preferably from Plasmodium faidparum isolate 3D7, and referred to herein as PfTMD and has the amino add sequence of SEQ ID NO: 77. In some embodiments, the TMD is heterologous. In some embodiments, the TMD is from HSVl-gD and referred to herein as HSV-1TMD and has the amino add sequence of SEQ ID NO:75.

[0785] In some embodiments, the Antigen comprises a Rh5 invasion complex polypeptide construct as defined above.

[0786] In some embodiments, the Antigen comprises a Rh5 polypeptide construct as defined above. In some embodiments, a Rh5 polypeptide construct is or comprises an Rh5 ordered domain construct as defined above. In some embodiments, a polyribonucleotide described herein has one of the following structures: cap-hAg-Kozak-Rh5 construct-FI-A30L70 cap-hAg-Kozak-sec- Rh5 construct -FI-A30L70 cap-hAg-Kozak- Rh5 construct -TMD-FI-A30L70 cap-hAg-Kozak-sec- Rh5 construct -TMD-FI-A30L70 cap-hAg-Kozak- Rh5 construct -Linker-TMD-FI-A30L70 cap-hAg-Kozak-sec- Rh5 construct - Linker-TMD-FI-A30L70 cap-hAg-Kozak- Rh5 construct -Multimerization-FI-A30L70 cap-hAg-Kozak-sec- Rh5 construct - Multimerization -FI-A30L70 cap-hAg-Kozak- Rh5 construct -Unker-Multimerlzation-FI-A30L70 cap-hAg-Kozak-sec- Rh5 construct -Llnker-Multimerizatlon -FI-A30L70 cap-hAg-Kozak- Rh5 construct -Self-assem-FI-A30L70 cap-hAg-Kozak-sec- Rh5 construct -Self-assem-FI-A30L70 cap-hAg-Kozak- Rh5 construct -Unker-Self-assem-Fl-A30L70 cap-hAg-Kozak-sec- Rh5 construct -Linker-Self-assem-FI-A30L70 cap-hAg-Kozak- Viralsec-Rh5 construct-FI-A30L70 cap-hAg-Kozak- HSV-1gDsec - Rh5 construct -FI-A30L70 cap-hAg-Kozak- Viralsec -Rh5 construct -TMD-FI-A30L70 cap-hAg-Kozak- HSV-1gDsec - Rh5 construct -TMD-FI-A30L70 cap-hAg-Kozak- Viralsec. -Rh5 construct -Unker-TMD-FI-A30L70 cap-hAg-Kozak- HSV-1gDsec - Rh5 construct - Linker-TMD-FI-A30L70 cap-hAg-Kozak- Viralsec -Rh5 construct - Multimerization-FI-A30L70 cap-hAg-Kozak- HSV-1gDsec - Rh5 construct - Multimerization -FI-A30L70 cap-hAg-Kozak- Viralsec - Rh5 construct -Unker-Multimerization-FI-A30L70 cap-hAg-Kozak- HSV-1gDsec - Rh5 construct -Unker-Multimerization -FI-A30L70 cap-hAg-Kozak- Viralsec -Rh5 construct -Self-assem-FI-A30L70 cap-hAg-Kozak- HSV-1gDsec - Rh5 construct -Self-assem-FI-A30L70 cap-hAg-Kozak- Viralsec -Rh5 construct -Linker-Self-assem-FI-A30L70 cap-hAg-Kozak- HSV-1gDsec - Rh5 construct -Linker-Self-assem-FI-A30L70

[0787] In some embodiments, the Antigen comprises a CyRPA polypeptide construct as defined above. In some embodiments, a polyribonucleotide described herein has one of the following structures: cap-hAg-Kozak- CyRPA construct-FI-A30L70 cap-hAg- Kozak-sec- CyRPA construct -FI-A30L70 cap-hAg-Kozak- CyRPA construct -TMD-FI-A30L70 cap-hAg-Kozak-sec- CyRPA construct -TMD-FI-A30L70 cap-hAg-Kozak- CyRPA construct -Linker-TMD-FI-A30L70 cap-hAg-Kozak-sec- CyRPA construct - Linker-TMD-FI-A30L70 cap-hAg-Kozak- CyRPA construct -Multlmerlzation-FI-A30L70 cap-hAg-Kozak-sec- CyRPA construct - Multimerization -FI-A30L70 cap-hAg-Kozak- CyRPA construct -Linker-Multimerization-FI-A30L70 cap-hAg-Kozak-sec- CyRPA construct -Linker-Multimerization -FI-A30L70 cap-hAg-Kozak- Virateec- CyRPA construct-FI-A30L70 cap-hAg-Kozak- HSV-1gDsec - CyRPA construct -FI-A30L70 cap-hAg-Kozak- Viralsec - CyRPA construct -TMD-FI-A30L70 cap-hAg-Kozak- HSV-1gDsec - CyRPA construct -TMD-FI-A30L70 cap-hAg-Kozak- Viralsec - CyRPA construct -Linker-TMD-FI-A30L70 cap-hAg-Kozak- HSV-1gDsec. - CyRPA construct - Linker-TMD-FI-A30L70 cap-hAg-Kozak- Viralsec - CyRPA construct - Multimerlzation-FI-A30L70 cap-hAg-Kozak- HSV-lgDsec - CyRPA construct - Multimerization -FI-A30L70 cap-hAg-Kozak- Viralsec - CyRPA construct -Linker-Multimerizatlon-Fl-A30L70 cap-hAg-Kozak- HSV-1gDsec. - CyRPA construct -Linker-Multimerization -FI-A30L70

[0788] In some embodiments, the Antigen comprises a P113 polypeptide construct as defined above. In some embodiments, a polyribonucleotide described herein has one of the following structures: cap-hAg-Kozak- P113 construct-FI-A30L70 cap-hAg-Kozak-sec- P113 construct -FI-A30L70 cap-hAg-Kozak- P113 construct -TMD-FI-A30L70 cap-hAg-Kozak-sec- P113 construct -TMD-FI-A30L70 cap-hAg-Kozak- P113 construct -Linker-TMD-FI-A30L70 cap-hAg-Kozak-sec- P113 construct - Unker-TMD-FI-A30L70 cap-hAg-Kozak- P113 construct -Multimerization-FI-A30L70 cap-hAg-Kozak-sec- P113 construct - Multimerization -FI-A30L70 cap-hAg-Kozak- P113 construct -Unker-Multimerization-FI-A30L70 cap-hAg-Kozak-sec- P113 construct -Linker-Miitimerization -FI-A30L70 cap-hAg-Kozak- Virateec- Pl 13 construct-FI-A30L70 cap-hAg-Kozak- HSV-lgDsec - P113 construct -FI-A30L70 cap-hAg-Kozak- Viralsec - P113 construct -TMD-FI-A30L70 cap-hAg-Kozak- HSV-1gDsec - P113 construct -TMD-FI-A30L70 cap-hAg-Kozak- Viralsec - P113 construct -Linker-TMD-FI-A30L70 cap-hAg-Kozak- HSV-lgDsec - P113 construct - Linker-TMD-FI-A30L70 cap-hAg-Kozak- Viralsec - P113 construct - Multimerization-FI-A30L70 cap-hAg-Kozak- HSV-1gDsec. - P113 construct - Muttimerization -FI-A30L70 cap-hAg-Kozak- Viralsec - P113 construct -Linker-Multimerization-FI-A30L70 cap-hAg-Kozak- HSV-1gDsec - P113 construct -Linker-Multimerization -FI-A30L70 [0789] In some embodiments, the different elements (sec, Antigen, TMD) of a Plasmodium polypeptide construct may be linked by one or more linkers, e.g., a linker selected from an amino add sequence as defined in Table 6. In some embodiments, a linker has the amino acid sequence GGSGGGGSGG (SEQ ID NO: 80). In some embodiments, a linker has the amino acid sequence GGGS (SEQ ID NO: 83). In some embodiments, a linker has the amino acid sequence GGGGSGGGGSGGGGS (SEQ ID NO: 84). In some embodiments, a linker has the amino acid sequence AGNRVRRSVG (SEQ ID NO: 85).

[0790] In some embodiments, the sequence encoding a Plasmodium polypeptide construct described herein comprises a modified nucleoside replacing (partially or completely, preferably completely) uridine, wherein the modified nudeoside is selected from the group consisting of pseudouridine (ip), Nl-methyl-pseudouridine (mlip), and 5-methyl-uridine.

[0791] In some embodiments, the sequence encoding a Plasmodium polypeptide construct described herein is codon-optimized.

[0792] In some embodiments, the G/C content of the sequence encoding a Plasmodium polypeptide construct described herein Is increased compared to the wiki type coding sequence.

[0793] In some embodiments, the RNA described herein comprises: a 5' UTR comprising the nudeotide sequence of SEQ ID NO: 111, or a nucleotide sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% Identity to the nudeotide sequence of SEQ ID NO: 111; a 3' UTR comprising the nudeotide sequence of SEQ ID NO: 117, or a nucleotide sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% Identity to the nucleotide sequence of SEQ ID NO: 117; and a poly-A sequence comprising the nucleotide sequence of SEQ ID NO: 114.

[0794] In some embodiments, the RNA described herein comprises: m27,3'-OGppp(ml2'-O) ApG as capping structure at the 5'-end of the RNA; a 5' UTR comprising the nucleotide sequence of SEQ ID NO: 111, or a nucleotide sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the nucleotide sequence of SEQ ID NO: 111; a 3' UTR comprising the nucleotide sequence of SEQ ID NO: 117, or a nucleotide sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the nucleotide sequence of SEQ ID NO: 117; and a poly-A sequence comprising the nucleotide sequence of SEQ ID NO: 114.

[0795] In some embodiments, the RNA is unmodified. In some embodiments, the RNA is modified. In some embodiments, the RNA comprises Nl-methyl-pseudouridine (m1Ψ ) in place of at least one uridine (e.g., in place of each uridine).

[0796] In some embodiments, the RNA described herein comprises: m27,3'-OGppp(ml2'-O) ApG as capping structure at the 5'-end of the RNA; a 5' UTR comprising the nudeodde sequence of SEQ ID NO: 111, or a nucleotide sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the nucleotide sequence of SEQ ID NO: 111; a 3' UTR comprising the nucleotide sequence of SEQ ID NO: 117, or a nucleotide sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the nucleotide sequence of SEQ ID NO: 117; and a poly-A sequence comprising the nucleotide sequence of SEQ ID NO: 114; and Nl-methyl-pseudouridine ( lip) in place of at least one uridine (e.g., in place of each uridine).

[0797] In some embodiments, RNA described herein (e.g., contained in the compositions/formulations of the present dsdosure and/or used in the methods of the present disclosure) may be presented as a product containing the vaccine RNA as active substance and other Ingredients comprising: ALC-0315 ((4- hydroxybutyl)azanediyl)bis(hexane-6,l-diyl)bis(2-hexyldecanoate), ALC-0159 (2-[(polyethylene glycol)-2000]-N,N- ditetradecylacetamide), l,2-Distearoyl-sn-glycero-3-phosphocho6ne (DSPC), and cholesterol.

[0798] In some embodiments, the RNA described herein is formulated or is to be formulated as a liquid, a sold, or a combination thereof.

[0799] In some embodiments, the RNA described herein is formulated or is to be formulated for injection. [0800] In some embodiments, the RNA described herein is formulated or is to be formulated for intramuscular administration.

[0801] In some embodiments, the RNA described herein Is formulated or is to be formulated as a composition, e.g., a pharmaceutical composition.

[0802] In some embodiments, the composition comprises a cationlcally Ionizable lipid. [0803] In some embodiments, the composition comprises a cationlcally Ionizable lipid and one or more additional lipids. In some embodiments, the one or more additional lipids are selected from polymer-conjugated lipids, neutral lipids, and combinations thereof. In some embodiments, the neutral lipids include phospholipids, steroid lipids, and combinations thereof. In some embodiments, the one or more additional lipids are a combination of a polymer-conjugated lipid, a phospholipid, and a steroid lipid.

[0804] In some embodiments, the composition comprises a cationlcally ionizable lipid; a polymer-conjugated lipid which Is a PEG-conjugated lipid; cholesterol; and a phospholipid. In some embodiments, the phospholipid is DSPC. In some embodiments, the phospholipid is DOPE.

[0805] In some embodiments, the composition comprises a cationlcally ionizable lipid; a polymer-conjugated lipid which is 2-[(polyethylene glycd)-2000]-N,N-ditetradecylacetamide; cholesterol; and a phospholipid. In some embodiments, the phospholipid is DSPC. In some embodiments, the phospholipid is DOPE.

[0806] In some embodiments, the composition comprises a cationlcally ionizable lipid which is ((4- hydroxybutyl)azanediyl)bis(hexane-6,l-diyl)bis(2-hexyldecanoate); a polymer-conjugated lipid which is 2-

[ (polyethylene glycol)-2000]-N,N-ditetradecylacetamide; cholesterol; and a phospholipid. In some embodiments, the phospholipid is DSPC. In some embedments, the phospholipid is DOPE In some embodiments, at least a portion of (i) the RNA, (ii) the cationlcally ionizable lipid, and if present, (iii) the one or more additional lipids is present in particles. In some embodiments, the particles are nanoparticles, such as lipid nanopartides (LNPs).

[0807] In some embodiments, the composition, in particular the pharmaceutical composition, is a vaccine.

[0808] In some embodiments, the composition, in particular the pharmaceutical composition, further comprises one or more pharmaceutically acceptable carriers, diluents and/or exdpients. [0809] In some embodiments, the RNA and/or the composition, in particular the pharmaceutical composition, is/are a component of a kit

[0810] In some embodiments, the kit further comprises instructions for use of the RNA for inducing an immune response against Plasmodium falciparum in a subject.

[0811] In some embodiments, the kit further comprises instructions for use of the RNA for therapeutically or prophylactically treating a Plasmodium falciparum infection in a subject.

[0812] In some embodiments, the subject is a human. [0813] In some embodiments, the RNA, e.g., RNA encoding a Plasmodium polypeptide construct, described in the present disclosure is non-immunogenlc. RNA encoding an Immunostimulant may be administered according to the present disclosure to provide an adjuvant effect The RNA encoding an Immunostimulant may be standard RNA or non-immunogenlc RNA.

E. Combinations of Polyribonucleotides

[0814] The present disclosure, among other things, utilizes RNA technologies as a modality to express two or more polypeptide constructs. In some embodiments, two or more different polypeptide constructs are two or more Plasmodium polypeptide constructs. As described further herein, in some embodiments, a Plasmodium polypeptide construct includes one or more malarial proteins, or one or more portions thereof.

[0815] In some embodiments, two or more Plasmodium polypeptide constructs indude a first Plasmodium polypeptide construct and a second Plasmodium polypeptide construct. In some embodiments, a first Plasmodium polypeptide construct comprises one or more Rh5 invasion complex polypeptide or antigenic portions thereof (e.g., a Rh5 polypeptide or antigenic portion thereof). In some embodiments, a first Plasmodium polypeptide construct comprises one or more Rh5 polypeptides or antigenic portions thereof. In some embodiments, a first Plasmodium polypeptide construct comprises one or more CyRPA polypeptides or antigenic portions thereof. In some embodiments, a first Plasmodium polypeptide construct comprises one or more P113 polypeptides or antigenic portions thereof. In some embodiments, a first Plasmodium polypeptide construct comprises one or more Ripr polypeptides or antigenic portions thereof. In some embodiments, a first Plasmodium polypeptide construct comprises one or more TRAMP polypeptides or antigenic portions thereof. In some embodiments, a first Plasmodium polypeptide construct comprises one or more CSS polypeptides or antigenic portions thereof.

[0816] In some embodiments, such a first Plasmodium polypeptide construct additionally includes one or more additional amino acid sequences, such as a secretory signal (e.g., a heterologous secretory signal), a transmembrane region (e.g., a heterologous transmembrane region), a multimerization region, a self-assembly region, and/or a linker, as described herein.

[0817] In some embodiments, a second Plasmodium polypeptide construct Is different than a first Hasmodium polypeptide construct. In some embodiments, a second Plasmodium polypeptide construct Includes one or more Plasmodium polypeptide regions or portions thereof (e.g., immunogenic fragments of a Plasmodium polypeptide), wherein the one or more Plasmodium polypeptide regions or portions thereof are different than one or more Plasmodium Rh5 invasion complex polypeptide regions or portions thereof included in a first Plasmodium polypeptide construct.

[0818] In some embodiments, a second Plasmodium polypeptide construct additionally Includes one or more additional amino add sequences, such as a secretory signal (e.g., a heterologous secretory signal), a transmembrane region (e.g., a heterologous transmembrane region), a multimerization region, a self-assembly region, and/or a linker, as described herein.

[0819] In some embodiments, a first Plasmodium polypeptide construct comprises one or more Plasmodium Rh5 polypeptides or antigenic portions thereof, and a second Plasmodium polypeptide construct includes one or more Plasmodium CyRPA polypeptides or antigenic portions thereof.

[0820] In some embodiments, a first Plasmodium polypeptide construct comprises one or more Plasmodium Rh5 polypeptides or antigenic portions thereof, and a second Plasmodium polypeptide construct includes one or more Plasmodium Pl 13 polypeptides or antigenic portions thereof.

[0821] In some embodiments, a first Plasmodium polypeptide construct comprises one or more Plasmodium Rh5 polypeptides or antigenic portions thereof, and a second Plasmodium polypeptide construct Includes one or more Plasmodium Ripr polypeptides or antigenic portions thereof.

[0822] In some embodiments, a first Plasmodium polypeptide construct comprises one or more Plasmodium Rh5 polypeptides or antigenic portions thereof, and a second Plasmodium polypeptide construct includes one or more Plasmodium TRAMP polypeptides or antigenic portions thereof.

[0823] In some embodiments, a first Plasmodium polypeptide construct comprises one or more Plasmodium Rh5 polypeptides or antigenic portions thereof, and a second Plasmodium polypeptide construct Includes one or more Plasmodium CSS polypeptides or antigenic portions thereof.

[0824] In some embodiments, a first Plasmodium polypeptide construct comprises one or more Plasmodium CyRPA polypeptides or antigenic portions thereof, and a second Plasmodium polypeptide construct includes one or more Plasmodium Rh5 polypeptides or antigenic portions thereof.

[0825] In some embodiments, a first Plasmodium polypeptide construct comprises one or more Plasmodium CyRPA polypeptides or antigenic portions thereof, and a second Plasmodium polypeptide construct includes one or more Plasmodium Pl 13 polypeptides or antigenic portions thereof.

[0826] In some embodiments, a first Plasmodium polypeptide construct comprises one or more Plasmodium CyRPA polypeptides or antigenic portions thereof, and a second Plasmodium polypeptide construct includes one or more Plasmodium Ripr polypeptides or antigenic portions thereof.

[0827] In some embodiments, a first Plasmodium polypeptide construct comprises one or more Plasmodium CyRPA polypeptides or antigenic portions thereof, and a second Plasmodium polypeptide construct includes one or more Plasmodium TRAMP polypeptides or antigenic portions thereof.

[0828] In some embodiments, a first Plasmodium polypeptide construct comprises one or more Plasmodium CyRPA polypeptides or antigenic portions thereof, and a second Plasmodium polypeptide construct includes one or more Plasmodium CSS polypeptides or antigenic portions thereof.

[0829] In some embodiments, a first Plasmodium polypeptide construct comprises one or more Plasmodium P113 polypeptides or antigenic portions thereof, and a second Plasmodium polypeptide construct includes one or more Plasmodium Rh5 polypeptides or antigenic portions thereof.

[0830] In some embodiments, a first Plasmodium polypeptide construct comprises one or more Plasmodium P113 polypeptides or antigenic portions thereof, and a second Plasmodium polypeptide construct includes one or more Plasmodium CyRPA polypeptides or antigenic portions thereof. [0831] In some embodiments, a first Plasmodium polypeptide construct comprises one or more Plasmodium P113 polypeptides or antigenic portions thereof, and a second Plasmodium polypeptide construct includes one or more Plasmodium Ripr polypeptides or antigenic portions thereof.

[0832] In some embodiments, a first Plasmodium polypeptide construct comprises one or more Plasmodium P113 polypeptides or antigenic portions thereof, and a second Plasmodium polypeptide construct includes one or more Plasmodium TRAMP polypeptides or antigenic portions thereof.

[0833] In some embodiments, a first Plasmodium polypeptide construct comprises one or more Plasmodium P113 polypeptides or antigenic portions thereof, and a second Plasmodium polypeptide construct includes one or more Plasmodium CSS polypeptides or antigenic portions thereof.

[0834] In some embodiments, a first Plasmodium polypeptide construct comprises one or more Plasmodium Ripr polypeptides or antigenic portions thereof, and a second Plasmodium polypeptide construct includes one or more Plasmodium Rh5 polypeptides or antigenic portions thereof.

[0835] In some embodiments, a first Plasmodium polypeptide construct comprises one or more Plasmodium Ripr polypeptides or antigenic portions thereof, and a second Plasmodium polypeptide construct includes one or more Plasmodium CyRPA polypeptides or antigenic portions thereof.

[0836] In some embodiments, a first Plasmodium polypeptide construct comprises one or more Plasmodium Ripr polypeptides or antigenic portions thereof, and a second Plasmodium polypeptide construct Includes one or more Plasmodium Pl 13 polypeptides or antigenic portions thereof.

[0837] In some embodiments, a first Plasmodium polypeptide construct comprises one or more Plasmodium Ripr polypeptides or antigenic portions thereof, and a second Plasmodium polypeptide construct includes one or more Plasmodium TRAMP polypeptides or antigenic portions thereof.

[0838] In some embodiments, a first Plasmodium polypeptide construct comprises one or more Plasmodium Ripr polypeptides or antigenic portions thereof, and a second Plasmodium polypeptide construct Includes one or more Plasmodium CSS polypeptides or antigenic portions thereof.

[0839] In some embodiments, a first Plasmodium polypeptide construct comprises one or more Plasmodium TRAMP polypeptides or antigenic portions thereof, and a second Plasmodium polypeptide construct includes one or more Plasmodium Rh5 polypeptides or antigenic portions thereof.

[0840] In some embodiments, a first Plasmodium polypeptide construct comprises one or more Plasmodium TRAMP polypeptides or antigenic portions thereof, and a second Plasmodium polypeptide construct includes one or more Plasmodium CyRP A polypeptides or antigenic portions thereof.

[0841] In some embodiments, a first Plasmodium polypeptide construct comprises one or more Plasmodium TRAMP polypeptides or antigenic portions thereof, and a second Plasmodium polypeptide construct includes one or more Plasmodium Ripr polypeptides or antigenic portions thereof.

[0842] In some embodiments, a first Plasmodium polypeptide construct comprises one or more Plasmodium TRAMP polypeptides or antigenic portions thereof, and a second Plasmodium polypeptide construct includes one or more Plasmodium Pl 13 polypeptides or antigenic portions thereof.

[0843] In some embodiments, a first Plasmodium polypeptide construct comprises one or more Plasmodium TRAMP polypeptides or antigenic portions thereof, and a second Plasmodium polypeptide construct includes one or more Plasmodium CSS polypeptides or antigenic portions thereof. [0844] In some embodiments, a first Plasmodium polypeptide construct comprises one or more Plasmodium CSS polypeptides or antigenic portions thereof, and a second Plasmodium polypeptide construct includes one or more Piasmocfium Rh5 polypeptides or antigenic portions thereof.

[0845] In some embodiments, a first Plasmodium polypeptide construct comprises one or more Plasmodium CSS polypeptides or antigenic portions thereof, and a second Plasmodium polypeptide construct includes one or more Plasmodium CyRPA polypeptides or antigenic portions thereof.

[0846] In some embodiments, a first Plasmodium polypeptide construct comprises one or more Plasmodium CSS polypeptides or antigenic portions thereof, and a second Plasmodium polypeptide construct includes one or more Plasmodium Ripr polypeptides or antigenic portions thereof.

[0847] In some embodiments, a first Plasmodium polypeptide construct comprises one or more Plasmodium CSS polypeptides or antigenic portions thereof, and a second Plasmodium polypeptide construct indudes one or more Plasmodium Pl 13 polypeptides or antigenic portions thereof.

[0848] In some embodiments, a first Plasmodium polypeptide construct comprises one or more Plasmodium CSS polypeptides or antigenic portions thereof, and a second Plasmodium polypeptide construct indudes one or more Plasmodium TRAMP polypeptides or antigenic portions thereof.

[0849] In some embodiments, a first Plasmodium polypeptide construct comprises one or more Plasmodium Rh5 Invasion complex polypeptide regions or portions thereof, and a second Plasmodium polypeptide construct indudes one or more Plasmodium polypeptide regions or portions thereof, wherein the one or more Plasmodium polypeptide regions or portions thereof comprise one or more Plasmodium CSP antigens.

[0850] In some embodiments, a first Plasmodium polypeptide construct comprises one or more Plasmodium Rh5 invasion complex antigens or portions thereof, and a second Plasmodium polypeptide construct Indudes one or more Plasmodium polypeptide regions or portions thereof, wherein the one or more Plasmodium polypeptide regions or portions thereof comprise one or more Plasmodium T-cell antigens.

[0851] In accordance with the above, the present disclosure provides two or more polyribonucleotides that each encode a polypeptide construct. In some embodiments, two or more polyribonucleotides that each encode a Plasmodium polypeptide construct. As described further herein, in some embodiments, a Plasmodium polypeptide construct Includes one or more malarial proteins, or one or more portions thereof.

[0852] In some embodiments, a first and second polyribonucleotide, as described herein, encode a Plasmodium polypeptide construct described herein has one of the following structures: cap-hAg-Kozak-Antigen-FI-A30L70 cap-hAg-Kozak-sec-Antigen-FI-A30L70 cap-hAg-Kozak-Antigen-TMD-FI-A30L70 cap-hAg-Kozak-sec-Antigen-TMD-FI-A30L70 cap-hAg-Kozak-Antigen-Linker-TMD-FI-A30L70 cap-hAg-Kozak-sec-Antigen- Unker-TMD-FI-A30L70 cap-hAg-Kozak-Antigen-Multimerization-FI-A30L70 cap-hAg-Kozak-sec-Antigen- Multimerization -FI-A30L70 cap-hAg-Kozak-Antjgen-Linker-Multimerization-FI-A30L70 cap-hAg-Kozak-sec-Antigen-Linker-Multimerization -FI-A30L70 cap-hAg-Kozak-Antigen-Self-assem-FI-A30L70 cap-hAg-Kozak-sec -Antigen-Self-assem-FI-A30L70 cap-hAg-Kozak-Antigen-Unker-Seif-assem -FI-A30L70 cap-hAg-Kozak-sec-Antigen-Linker-Self assem-FI-A30L70

[0853] In some embodiments, hAg-Kozak comprises the nucleotide sequence of SEQ ID NO: 111. In some embodiments, FI comprises the nudeotide sequence of SEQ ID NO: 117. In some embodiments, A30L70 comprises the nucleotide sequence of SEQ ID NO: 114.

[0854] In some embodiments, the secretory signal is from Plasmodium faidparum, preferably Plasmodium falciparum isolate 3D7. In some embodiments, a secretory signal is from a Rh5 invasion complex polypeptide of Plasmodium faidparum (e.g., Isolate 3D7). In some embodiments, a secretory signal is from a Rh5 polypeptide of Plasmodium faidparum. In some embodiments, a secretory signal Is from a CSP polypeptide of Plasmodium falciparum and has the sequence as defined in Table 3 (SEQ ID NO: 41). In some embodiments, the secretory signal is heterologous and selected from the amino acid sequences as defined in Table 3. In some embodiments, the secretory signal is HSVl-gD and referred to herein as HSV-1gDsec and has the sequence as defined In Table 3 (SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 42).

[0855] In some embodiments, the TMD Is a glycosylphosphatidyHnosItoi (GPI) anchor region from Plasmodium CSP, preferably from Plasmodium faidparum isolate 3D7, and referred to herein as 77TMD and has the amino add sequence of SEQ ID NO: 77. In some embodiments, the TMD Is heterologous. In some embodiments, the TMD Is from HSVl-gD and referred to herein as /75V-1TMD and has the amino add sequence of SEQ ID NO:75.

[0856] In some embodiments, the different elements (sec, Antigen, TMD) of a first polypeptide may be Hnked by one or more linkers, e.g., a linker selected from an amino add sequence as defined In Table 6. In some embodiments, a linker has the amino acid sequence GGSGGGGSGG (SEQ ID NO: 80). In some embodiments, a linker has the amino acid sequence GGGS (SEQ ID NO: 83). In some embodiments, a linker has the amino add sequence GGGGSGGGGSGGGGS (SEQ ID NO: 84). In some embodiments, a linker has the amino acid sequence AGNRVRRSVG (SEQ ID NO: 85).

[0857] In some embodiments, a combination as provided herein can indude (I) a first polyribonucleotide, where the first polyribonucleotide encodes a first polypeptide; and (II) a second polyribonucleotide, where the second polyribonucleotide encodes a second polypeptide, and the second polypeptide comprises one or more Plasmodium polypeptide regions or portions thereof.

[0858] In some embodiments, the first polyribonucleotide has one of the following structures: cap-hAg-Kazak-Rh5 construct-FI-A30L70 cap-hAg-Kozak-sec- Rh5 construct -FI-A30L70 cap-hAg-Kozak- Rh5 construct -TMD-FI-A30L70 cap-hAg-Kozak-sec- Rh5 construct -TMD-FI-A30L70 cap-hAg-Kozak- Rh5 construct -Linker-TMD-FI-A30L70 cap-hAg-Kozak-sec- Rh5 construct - Linker-TMD-FI-A30L70 cap-hAg-Kozak- Rh5 construct -Multimerization-FI-A30L70 cap-hAg-Kozak-sec- Rh5 construct - Multimerization -FI-A30L70 cap-hAg-Kozak- Rh5 construct -Unker-Multimerization-FI-A30L70 cap-hAg-Kozak-sec- Rh5 construct -Linker-Multimerization -FI-A30L70 cap-hAg-Kozak- Rh5 construct -Self-assem-FI-A30L70 cap-hAg-Kozak-sec- Rh5 construct -Sdf-assem-FI-A30L70 cap-hAg-Kozak- Rh5 construct -Unker-Sdf-assem-FI-A30L70 cap-hAg-Kozak-sec- Rh5 construct -Linker-Self-assem-FI-A30L70 cap-hAg-Kozak- Viralsec-Rh5 construct-FI-A30L70 cap-hAg-Kozak- HSV-1gDsec - Rh5 construct -FI-A30L70 cap-hAg-Kozak- Viralsec -Rh5 construct -TMD-FI-A30L70 cap-hAg-Kozak- HSV-1gDsec - Rh5 construct -TMD-FI-A30L70 cap-hAg-Kozak- Viralsec -Rh5 construct -Linker-TMD-FI-A30L70 cap-hAg-Kozak- HSV-1gDsec - Rh5 construct - Linker-TMD-FI-A30L70 cap-hAg-Kozak- Virafeec -Rh5 construct - Multlmerization-FI-A30L70 cap-hAg-Kozak- HSV-1gDsec - Rh5 construct - Multimerization -FI-A30L70 cap-hAg-Kozak- Viralsec - Rh5 construct -Linker-Multimerization-FI-A30L70 cap-hAg-Kozak- HSV-1gDsec. - Rh5 construct -Linker-Multimerization -FI-A30L70 cap-hAg-Kozak- Viralsec -Rh5 construct -Self-assem-FI-A30L70 cap-hAg-Kozak- HSV-1gDsec - Rh5 construct -Self-assem-FI-A30L70 cap-hAg-Kdzak- Viralsec -Rh5 construct -Llnker-Self-assem-FI-A30L70 cap-hAg-Kozak- HSV-1gDsec - Rh5 construct -Unker-Self-assem-FI-A30L70

[0859] In some embodiments, the Antigen comprises a CyRPA polypeptide construct as defined above. In some embodiments, a polyribonucleotide described herein has one of the following structures: cap-hAg-Kozak- CyRPA construct-FI-A30L70 cap-hAg-Kozak-sec- CyRPA construct -FJ-A30L70 cap-hAg-Kozak- CyRPA construct -TMD-FI-A30L70 cap-hAg-Kozak-sec- CyRPA construct -TMD-FI-A30L70 cap-hAg-Kozak- CyRPA construct -Linker-TMD-FI-A30L70 cap-hAg-Kozak-sec- CyRPA construct - Linker-TMD-FI-A30L70 cap-hAg-Kozak- CyRPA construct -Multimerization-FI-A30L70 cap-hAg-Kozak-sec- CyRPA construct - Multimerization -FI-A30L70 cap-hAg-Kozak- CyRPA construct -Linker-Multimerization-FI-A30L70 cap-hAg-Kozak-sec- CyRPA construct -Linker-Multimerization -FI-A30L70 cap-hAg-Kozak- Viralsec- CyRPA construct-FI-A30L70 cap-hAg-Kozak- HSV-lgDsec - CyRPA construct -FI-A30L70 cap-hAg-Kozak- Viralsec - CyRPA construct -TMD-FI-A30L70 cap-hAg-Kozak- HSV-1gDsec - CyRPA construct -TMD-FI-A30L70 cap-hAg-Kozak- Viralsec - CyRPA construct -Linker-TMD-FI-A30L70 cap-hAg-Kozak- HSV-1gDsec - CyRPA construct - Linker-TMD-FI-A30L70 cap-hAg-Kozak- Viralsec - CyRPA construct - Multimerization-FI-A30L70 cap-hAg-Kozak- HSV-1gDsec - CyRPA construct - Multimerization -FI-A30L70 cap-hAg-Kozak- Viralsec - CyRPA construct -Linker-Multimerization-FI-A30L70 cap-hAg-Kozak- HSV-1gDsec - CyRPA construct -Linker-Multimerization -FI-A30L70 [0860] In some embodiments, the Antigen comprises a P113 polypeptide construct as defined above. In some embodiments, a polyribonucleotide described herein has one of the following structures: cap-hAg-Kozak- P113 construct-FI-A30L70 cap-hAg-Kozak-sec- P113 construct -FI-A30L70 cap-hAg-Kozak- P113 construct -TMD-FI-A30L70 cap-hAg-Kozak-sec- P113 construct -TMD-FI-A30L70 cap-hAg-Kozak- P113 construct -Linker-TMD-FI-A30L70 cap-hAg-Kozak-sec- P113 construct - Linker-TMD-FI-A30L70 cap-hAg-Kozak- P113 construct -Multimerizatlon-FI-A30L70 cap-hAg-Kozak-sec- P113 construct - Multimerizatlon -FI-A30L70 cap-hAg-Kozak- P113 construct -Unker-Multimerization-FI-A30L70 cap-hAg-Kozak-sec- P113 construct -Unker-Mdtimerization -FI-A30L70 cap-hAg-Kozak- Viralsec - Pl 13 construct-FI-A30L70 cap-hAg-Kozak- HSV-1gDsec - P113 construct -FI-A30L70 cap-hAg-Kozak- Viralsec - P113 construct -TMD-FI-A30L70 cap-hAg-Kozak- HSV-1gDsec - P113 construct -TMD-FI-A30L70 cap-hAg-Kozak- Viralsec - P113 construct -Unker-TMD-FI-A30L70 cap-hAg-Kozak- HSV-1gDsec. - P113 construct - Linker-TMD-FI-A30L70 cap-hAg-Kozak- Viralsec - P113 construct - Multimerization-FI-A30L70 cap-hAg-Kozak- HSV-1gDsec. - P113 construct - Multimerizatlon -FI-A30L70 cap-hAg-Kozak- Vfrafeec - P113 construct -Unker-Multimerization-FI-A30L70 cap-hAg-Kozak- HSV-1gDsec - P113 construct -Unker-Multimerization -FI-A30L70 [0861] In some embodiments, a first and/or a second polyribonucleotide described herein comprises a modified nucleoside replacing (partially or completely, preferably completely) uridine, wherein the modified nucleoside is selected from the group consisting of pseudouridine (Ψ ), Nl-methyl-pseudourkfine (mlΨ), and 5- methyl-uridlne.

[0862] In some embodiments, a first and/or a second polyribonucleotide described herein Is codon-optimized. [0863] In some embodiments, the G/C content of a first and/or a second polyribonucleotide described herein is increased compared to the wild type coding sequence.

[0864] In some embodiments, a first and/or a second polyribonucleotide described herein comprises: a 5' UTR comprising the nucleotide sequence of SEQ ID NO: 111, or a nucleotide sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% Identity to the nucleotide sequence of SEQ ID NO: 111; a 3' UTR comprising the nucleotide sequence of SEQ ID NO: 117, or a nucleotide sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the nucleotide sequence of SEQ ID NO: 117; and a poty-A sequence comprising the nucleotide sequence of SEQ ID NO: 114.

[0865] In some embodiments, a first and/or a second polyribonucleotide described herein comprises: m27,3'-OGppp(ml2'-O) ApG as capping structure at the 5'-end of the RNA; a 5' UTR comprising the nucleotide sequence of SEQ ID NO: 111, or a nucleotide sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the nucleotide sequence of SEQ ID NO: 111; a 3' UTR comprising the nucleotide sequence of SEQ ID NO: 117, or a nucleotide sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the nucleotide sequence of SEQ ID NO: 117; and a poly-A sequence comprising the nucleotide sequence of SEQ ID NO: 114.

[0866] In some embodiments, a first and/or a second polyribonucleotide described herein is unmodified. In some embodiments, a first and/or a second polyribonucleotide described herein is modified. In some embodiments, a first and/or a second polyribonucleotide described herein comprises Nl-methyl-pseudouridine (mini) in place of at least one uridine (e.g., in place of each uridine).

[0867] In some embodiments, a first and/or a second polyribonucleotide described herein comprises: m27,3'-OGppp(ml2'-O) ApG as capping structure at the 5 -end of the RNA; a 5' UTR comprising the nucleotide sequence of SEQ ID NO: 111, or a nucleotide sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the nucleotide sequence of SEQ ID NO: 111; a 3' UTR comprising the nucleotide sequence of SEQ ID NO: 117, or a nucleotide sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% Identity to the nucleotide sequence of SEQ ID NO: 117; and a poiy-A sequence comprising the nucleotide sequence of SEQ ID NO: 114, and Nl-methyl-pseudourldlne (mlΨ ) In place of at least one uridine (e.g., in place of each uridine).

[0868] In some embodiments, a combination described above can be administered in a pharmaceutical composition as described herein. In some embodiments, two or more polyribonucleotides of a combination as described above can be administered In separate pharmaceutical compositions as described herein. For example, in some embodiments, a combination provided herein comprises (I) a first pharmaceutical composition comprising a first polyribonucleotide, wherein the first polyribonucleotide encodes a first polypeptide, and the first polypeptide comprises one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof; and (ii) a second pharmaceutical composition comprising a second polyribonucleotide, wherein the second polyribonucleotide encodes a second polypeptide, and the second polypeptide comprises one or more polypeptides or antigenic portions thereof.

[0869] In some embodiments, a first or a second polyribonucleotide as described herein (e.g., contained in the compositions/formulations of the present disclosure and/or used in the methods of the present disclosure) may be presented as a product containing the first or the second polyribonucleotide as described herein as active substance and other ingredients comprising: ALC-0315 ((4-hydroxybutyl)azanediyl)bis(hexane-6,l-diyl)bis(2-hexyldecanoate), ALC-0159 (2-[(polyethylene glycol)-2000]-N,N-ditetradecylacetamide), l,2-Distearoyl-sn-glycero-3-phosphocholine (DSPC), and cholesterol.

[0870] In some embodiments, a first or a second polyribonucleotide as described herein is formulated or is to be formulated as a liquid, a solid, or a combination thereof.

[0871] In some embodiments, a first or a second polyribonucleotide as described herein is formulated or is to be formulated for injection.

[0872] In some embodiments, a first or a second polyribonucleotide as described herein is formulated or is to be formulated for intramuscular administration.

[0873] In some embodiments, a first or a second polyribonucleotide as described herein is formulated or Is to be formulated as a composition, e.g., a pharmaceutical composition. [0874] In some embodiments, a composition comprises a cationically ionizable lipid. [0875] In some embodiments, a composition comprises a cationically ionizable lipid and one or more additional lipids. In some embodiments, one or more additional lipids are selected from pdymer-conjugated lipids, neutral lipids, and combinations thereof. In some embodiments, neutral lipids include phospholipids, steroid lipids, and combinations thereof. In some embodiments, one or more additional lipids are a combination of a pdymer- conjugated lipid, a phospholipid, and a steroid lipid.

[0876] In some embodiments, a composition comprises a cationically ionizable lipid; a pdymer-conjugated lipid which is a PEG-conjugated lipid; cholesterol; and a phosphdipid. In some embodiments, a phosphdipid is DSPC. In some embodiments, a phosphdipid is DOPE.

[0877] In some embodiments, a composition comprises a cationically ionizable lipid; a pdymer-conjugated lipid which is 2-[(pdyethylene glycol)-2000]-N,N-ditetradecyiacetamide; cholesterol; and a phospholipid. In some embodiments, a phospholipid is DSPC. In some embodiments, a phosphdipid is DOPE.

[0878] In some embedments, a composition comprises a cationically ionizable lipid which is ((4- hydroxybutyl)azanediyl)bis(hexane-6,l-diyl)bis(2-hexyldecanoate); a pdymer-conjugated lipid which is 2- [ (polyethylene glycd)-2000]-N,N-ditetradecylacetamide; cholesterol; and a phosphdipid. In some embodiments, a phosphdipid is DSPC. In some embodiments, a phosphdipid is DOPE. In some embodiments, at least a portion of (I) a first or a second polyribonucleotide as described herein, (ii) a cationically Ionizable lipid, and if present, (III) one or more additional lipids is present in particles. In some embodiments, particles are nanoparticles, such as lipid nanoparticles (LNPs).

[0879] In some embodiments, a composition, In particular the pharmaceutical composition, Is a vaccine. [0880] In some embodiments, a composition, in particular the pharmaceutical composition, further comprises one or more pharmaceutically acceptable carriers, diluents and/or excipients.

IV. RNA Delivery Technologies

[0881] Provided polyribonucleotides may be delivered for therapeutic applications described herein using any appropriate methods known in the art, including, e.g., deBvery as naked RNAs, or delivery mediated by viral and/or non-vlral vectors, polymer-based vectors, lipid compositions, nanopartides ^e.g., lipid nanoparticles, polymeric nanopartides, llpld-pdymer hybrid nanoparticles, etc.), and/or peptide-based vectors. See, e.g., Wadhwa etai.

"Opportunities and Challenges in the Delivery of mRNA-Based Vaccines" Pharmaceutics (2020) 102 (27 pages), the content of which is incorporated herein by reference, for information on various approaches that may be useful for delivery polyribonucleotides described herein.

[0882] In some embodiments, one or more polyribonucleotides can be formulated with lipid nanopartides for delivery ^e.g., administration).

[0883] In some embodiments, lipid nanopartides can be designed to protect polyribonucleotides from extracellular RNases and/or engineered for systemic deBvery of the RNA to target cells (e.g., liver cells). In some embodiments, such lipid nanopartides may be particularly useful to deliver polyribonucleotides when pdyribonudeotides are intravenously or intramuscularly administered to a subject.

Lipid Compositions

1. Lipids and Lipid-Like Materials

[0884] The terms "lipid" and "lipid-like material" are broadly defined herein as molecules which comprise one or more hydrophobic moieties or groups and optionally also one or more hydrophilic moieties or groups. Molecules comprising hydrophobic moieties and hydrophilic moieties are also frequently denoted as amphiphiles. Lipids are usually poorly soluble in water. In an aqueous environment, the amphiphilic nature allows the molecules to selfassemble into organized structures and different phases. One of those phases consists of lipid bilayers, as they are present in vesicles, muftilamellar/unilamellar liposomes, or membranes in an aqueous environment. Hydrophobicity can be conferred by the inclusion of a polar groups that indude, but are not limited to, long-chain saturated and unsaturated aliphatic hydrocarbon groups and such groups substituted by one or more aromatic, cycloaliphatic, or heterocydic group(s). The hydrophilic groups may comprise polar and/or charged groups and include carbohydrates, phosphate, carboxylic, sulfate, amino, sulfhydryl, nitro, hydroxyl, and other like groups.

[0885] Often, an amphiphilic compound has a polar head attached to a long hydrophobic tail. In some embodiments, the polar portion Is soluble in water, while the non-polar portion Is insoluble In water. In addition, the polar portion may have either a formal positive charge, or a formal negative charge. Alternatively, the polar portion may have both a formal positive and a negative charge, and be a zwitterion or inner salt. For purposes of the disdosure, the amphiphile compound can be, but Is not limited to, one or a plurality of natural or non-natural lipids and Hpid-like compounds.

[0886] A "lipid-like material" is a substance that is structurally and/or functionally related to a lipid but may not be considered a lipid in a strict sense. For example, the term Indudes compounds that are able to form amphiphilic layers as they are present In vesicles, multilamellar/unllamelar liposomes, or membranes In an aqueous environment and Indudes surfactants, or synthesized compounds with both hydrophilic and hydrophobic moieties. Generally speaking, the term refers to molecules, which comprise hydrophilic and hydrophobic moieties with different structural organization, which may or may not be similar to that of lipids.

[0887] Specific examples of amphiphilic compounds that may be Induded In an amphiphilic layer indude, but are not limited to, phospholipids, aminolipids and sphingolipids.

[0888] Generally, lipids may be divided Into eight categories: fatty adds, glycerollpids, glycerophospholipids, sphingolipids, saccharollplds, poiyketides (derived from condensation of ketoacyl subunits), sterols and prenol lipids (derived from condensation of isoprene subunits). Although the term "Ipki" Is sometimes used as a synonym for fats, fats are a subgroup of lipids called triglycerides. Lipids also encompass molecules such as fatty adds and their derivatives (indixfing tri-, di-, monoglycerides, and phospholipids), as well as sterol-contalnlng metabolites such as cholesterol.

[0889] Fatty adds are a diverse group of molecules made of a hydrocarbon chain that terminates with a carboxylic add group; this arrangement confers the molecule with a polar, hydrophilic end, and a nonpolar, hydrophobic end that is insoluble in water. "The carbon chain, typically between four and 24 carbons long, may be saturated or unsaturated, and may be attached to functional groups containing oxygen, halogens, nitrogen, and sulfur. If a fatty acid contains a double bond, there is the possibility of either a ds or trans geometric isomerism, which significantly affects the molecule's configuration. Os-double bonds cause the fatty acid chain to bend, an effect that is compounded with more double bonds in the chain. Other major lipid classes in the fatty add category are the fatty esters and fatty amides.

[0890] Glycerollpids are composed of mono-, di-, and tri-substituted glycerols, the best-known being the fatty add triesters of glycerol, called triglycerides. The word "triacylglyceror is sometimes used synonymously with "triglyceride". In these compounds, the three hydroxyl groups of glycerol are each esterified, typically by different fatty acids. Additional subclasses of glycerolipids are represented by glycosylglycerols, which are characterized by the presence of one or more sugar residues attached to glycerol via a glycosidic linkage.

[0891] Glycerophospholipids are amphipathic molecules (containing both hydrophobic and hydrophilic regions) that contain a glycerol core linked to two fatty add-derived "tails" by ester linkages and to one "head" group by a phosphate ester linkage. Examples of glycerophospholipids, usually referred to as phospholipids (though sphingomyelins are also classified as phospholipids) are phosphatidylcholine (also known as PC, GPCho or lecithin), phosphatidylethanolamine (PE or GPEtn) and phosphatidylserine (PS or GPSer).

[0892] Sphingolipids are members of a complex family of compounds that share a common structural feature, a sphingoid base backbone. "The major sphingoid base in mammals Is commonly referred to as sphingosine.

Ceramides (N-acyl-sphingold bases) are a major subclass of sphingoid base derivatives with an amide-linked fatty acid. The fatty adds are typically saturated or mono-unsaturated with chain lengths from 16 to 26 carbon atoms. The major phosphosphingolipids of mammals are sphingomyeins (ceramide phosphocholines), whereas insects contain mainly ceramide phosphoethanolamines and fungi have phytoceramide phosphoinositols and mannose-containing headgroups. The glycosphingolipids are a diverse family of molecules composed of one or more sugar residues linked via a glycosidic bond to the sphingoid base. Examples of these are the simple and complex glycosphingolipids such as cerebrosides and gangliosides.

[0893] Sterols, such as cholesterol and Its derivatives, or tocopherol and Its derivatives, are Important components of membrane lipids, along with the glycerophospholipids and sphingomyelins.

[0894] Saccharolipids are compounds in which fatty adds are linked directly to a sugar backbone, forming structures that are compatible with membrane bllayers. In the saccharolipids, a monosaccharide substitutes for the glycerol backbone present in glycerolipids and glycerophospholipids. The most familiar saccharolipids are the acylated glucosamine precursors of the Lipid A component of the lipopolysaccharides in Gram-negative bacteria. Typical lipid A molecules are disaccharides of glucosamine, which are derivatized with as many as seven fatty-acyi chains. The minimal lipopolysaccharide required for growth in E. coll is Kdo2 -Lipid A, a hexa-acylated disaccharide of glucosamine that Is glycosylated with two 3-deoxy-D-manno-octuk)sonlc add (Kdo) residues.

[0895] Polyketldes are synthesized by polymerization of acetyl and propionyl subunits by classic enzymes as well as iterative and multimodiiar enzymes that share mechanistic features with the fatty add synthases. They comprise a large number of secondary metabolites and natural products from animal, plant, bacterial, fungal and marine sources, and have great structural diversity. Many polyketldes are cydic molecules whose backbones are often further modified by glycosylation, methylation, hydroxylation, oxidation, or other processes.

[0896] Lipids and ipid-like materials may be cationic, anionic or neutral. Neutral lipids or lipid-like materials exist In an uncharged or neutral zwitterionic form at a selected pH.

[0897] In some embodiments, suitable lipids or lipid-like materials for use in the present disclosure indude those described in W02020/128031 and US20200163878, the entire contents of each of which are incorporated herein by reference for the purposes described herein.

2. Cationic or cationicaily ionizable lipids or Kpid-Hke materials

[0898] In some embodiments cationic or cationicaily ionizable lipids or ipid-like materials contemplated for use herein include any cationic or cationicaily ionizable lipids or ipid-like materials which are able to electrostatically bind nudeic add. In one embodiment, cationic or cationicaily ionizable lipids or lipid-like materials contemplated for use herein can be associated with nucleic acid, e.g. by forming complexes with the nucleic add or forming vesicles in which the nucleic add is endosed or encapsulated.

[0899] Cationic ipids or lipid-like materials are characterized in that they have a net positive charge (e.g., at a relevant pH). Cationic lipids or lipid-like materials bind negatively charged nucleic acid by electrostatic interaction. Generally, cationic lipids possess a lipophilic moiety, such as a sterol, an acyl chain, a diacyl or more acyl chains, and the head group of the lipid typically carries the positive charge.

[0900] In certain embodiments, a cationic lipid or lipid-like material has a net positive charge only at certain pH, in particular addic pH, while it has preferably no net positive charge, preferably has no charge, i.e., it is neutral, at a different, preferably higher pH such as physiological pH. This ionizable behavior is thought to enhance efficacy through helping with endosomal escape and reducing toxldty as compared with particles that remain cationic at physiological pH.

[0901] In some embodiments, a cationic or cationically ionizable lipid or lipid-like material comprises a head group which indudes at least one nitrogen atom (N) which is positive charged or capable of being protonated. [0902] Examples of cationic lipids include, but are not limited to l,2-dloleoyl-3-trimethylammonium propane (DOTAP); N,N-dimethyl-2,3-dioleyloxypropylamine (DODMA), l,2-di-O-octadecenyl-3-trimethylammonium propane (DOTMA), 3-(N— (N',N'-dimethylaminoethane)-carbamoyl)cholesterol (DC -Choi), dimethytdioctadecylammonlum (DDAB); l,2-dk>leoyl-3-dimethylammonlum-propane (DODAP); l,2-dlacyk)xy-3-dimethylammonium propanes; 1,2- dialkyloxy-3-dimethylammonlum propanes; dioctadecykiimethyl ammonium chloride (DODAC), l,2-distearyioxy-N,N- dimethyl-3-aminopropane (DSDMA), 2,3-dl(tetradecoxy)propyl-(2-hydroxyethyl)-dlmethylazanlum (DMRIE), 1,2- dimyristoyl-sn-glycero-3-ethylphosphocholine (DMEPQ, l,2-dimyristoyl-3-trimethylammonlum propane (DMTAP), 1,2- dioleyloxypropyl-3-dimethyl-hydroxyethyl ammonium bromide (DORIE), and 2,3-dioleoyloxy- N-[2(spermine carboxamide)ethyl]-N,N-dimethyl-l-propanamium trifluoroacetate (DOSPA), l,2-dllinoleyloxy-N,N- dimethylamlnopropane (DLinDMA), l,2-dilinolenytoxy-N,N-dlmethylaminopropane (DLenDMA), dioctadecylamidoglycyl spermine (DOGS), 3-dlmethylamino-2-(cholest-5-en-3-beta-oxybutan-4-oxy)-l-(cis,ds-9,12-oc- tadecadlenoxy)propane (CUnDMA), 2-[5'-(cholest-5-en-3-beta-oxy)-3'-oxapentoxy)-3-dlmethyl-l-(cis,ds-9',12'- octadecadlenoxy)propane (CpLinDMA), N,N-dimethyl-3,4-dloleyloxybenzylamlne (DMOBA), l,2-N,N'-dioleylcarbamyl- 3-dimethylaminopropane (DOcarbDAP), 2,3-Dilinoleoyloxy-N,N-dimethylpropylamine (DUnDAP), 1,2-N,N'- Dilinoleyicarbamyi-3-dimethylaminopropane (DUncarbDAP), l,2-Dilinoleoylcarbamyl-3-dimethylaminopropane (DLinCDAP), 2,2-dilinoleyl-4-dimethylaminomethyl-[l,3]-dioxolane (DLin-K-DMA), 2,2-dilinoleyl-4-dimethylaminoethyl- [l,3]-dioxolane (DLin-K-XTC2-DMA), 2,2-dilinoleyl-4-(2-dimethylaminoethyl)-[l,3]-dioxoiane (DUn-KC2-DMA), heptatriaconta-6,9,28,31-tetraen-19-yl-4-(dimethylamino)butanoate (DLin-MC3-DMA), N-(2-Hydroxyethyl)-N,N- dimethyl-2,3-bis(tetradecyloxy)-l-propanamirtum bromide (DMRIE), (±)-N-(3-amlnopropyl)-N,N-dimethyl-2,3-bis(cis- 9-tetradecenytoxy)-l-propanaminium bromide (GAP-DMORIE), (±)-N-(3-aminopropyl)-N,N-dimethyl-2,3- bis(dodecyloxy)-l-propanaminium bromide (GAP-DLRIE), (±)-N-(3-aminopropyl)-N,N-dimethyl-2,3- bis(tetradecyloxy)-l-propanaminium bromide (GAP-DMRIE), N-(2-Aminoethyi)-N,N-dimethyl-2,3-bis(tetradecyloxy)-l- propanaminium bromide (gAE-DMRIE), N-(4-carboxybenzyl)-N,N-dimethyl-2,3-bis(oleoyloxy)propan-l-aminium (DOBAQ), 2-({8-[(3P)-cholest-5-en-3-yloxy]octyl}oxy)-N,N-dimethyl-3-[(9Z,12Z)-octadeca-9,12-dien-l-yloxy]propan- 1 -amine (Octyl-CLinDMA), l,2-dimyristoy1-3-dimethyiammonium-propane (DMDAP), l,2-dipalmitoyl-3- dimethylammonium-propane (DPDAP), Nl-[2-((lS)-l-[(3-amlnopropyl)amino]-4-[di(3-amino- propyl)amino]butylcarboxamido)ethyl]-3,4-di[oleyloxy]-benzamide (MVL5), l,2-dioleoyl-sn-glycero-3- ethylphosphocholine (DOEPC), 2,3-bis(dodecyloxy)-N-(2-hydroxyethyl)-N,N-dimethylpropan-l-amonium bromide (DLRIE), N-(2-aminoethyl)-N,N-dimethyl-2,3-bis(tetradecyloxy)propan-l-aminium bromide (DMORIE), di((Z)-non-2- en-l-yl) 8,8'-((((2(dimethylamino)ethyl)thio)carbonyl)azanediyl)dioctanoate (ATX), N,N-dimethyl-2,3- bis(dodecyloxy)propan-l-amine (DLDMA), N,N-dimethyl-2,3-bis(tetradecyloxy)propan-l-amine (DMDMA), Di((Z)-non- 2-en-l-yl)-9-((4-(dimethylaminobutanoyi)oxy)heptadecane<fioate (L319), N-Dodecyl-3-((2-dodecylcarbamoyl-ethyl)- {2-[(2-dodecylcarbamoyl-ethyl)-2-{(2-dodecylcart)amoyl-ethyl)-[2-(2-dodecylcarbamoyl-ethylamirK))-ethyl]-amino}- ethylamino)propionamide (lipidoid 98N12-5), l-[2-[bis(2-hydroxydodecyi)amino]ethyl-[2-[4-[2-[bis(2 hydroxydodecyl)amino]ethyl]piperazin-l-yl]ethyl]amino]dodecan-2-ol (lipidoid C12-200), LIPOFECTIN® (commercially available cationic liposomes comprising DOTMA and 1 ,2-dioleoyl-sn-3phosphoethanoiamine (DOPE), from GIBCO/BRL, Grand Island, N.Y.); LIPOFECTAMINE® (commercially available cationic liposomes comprising N-(l -(2,3dioleyloxy)propyl)-N-(2-(sperminecarboxamido)ethyl)-N,N-dimethylammonium trifluoroacetate (DOSPA) and (DOPE), from GIBCO/BRL); and TRANSFECTAM® (commercially available cationic lipids comprising dioctadecylamidoglycyl carboxyspermine (DOGS) in ethanol from Promega Corp., Madison, Wis.) or any combination of any of the foregoing. Further suitable cationic lipids for use in the present disclosure include those described in W02020/128031 and US20200163878, the entire contents of each of which are incorporated herein by reference for the purposes described herein. Further suitable cationic lipids for use in the present disclosure include those described In W02010/053572 (Including d 2-200 deserted at paragraph [00225]) and W02012/170930, both of which are incorporated herein by reference for the purposes described herein. Additional suitable cationic lipids for use In the present disclosure Include HGT4003, HGT5000, HGTS001, HGT5001, HGT5002 (see US20150140070A1, which is Incorporated herein by reference in its entirety).

[0903] In some embodiments, formulations that are useful for pharmaceutical compositions (e.g., Immunogenic compositions, e.g., vaccines) compositions as described herein can comprise at least one cationic lipid. Representative cationic lipids Include, but are not limited to, 1 ,2-dilinoleyoxy-3-(dimethylamino)acetoxypropane (DLin-DAC), 1 ,2-dilinoleyoxy-3morphollnopropane (DLln-MA), l,2-diHnoleoyl-3-dlmethylaminopropane (DUnDAP), 1 ,2-diinoleylthlo-3-dimethylaminopropane (DUn-S-DMA), 1 -linoleoyl-2-Hnoleyloxy-3dimethylaminopropane (DLIn-2- DMAP), 1 ,2-dilinoleyloxy-3-frimethylaminopropane chloride salt (DUn-TMA.CJ), 1 ,2-dilinoleoyl-3- trimethylaminopropane chloride salt (DUn-TAP.CI), 1 ,2-dilinoleyloxy-3-(N-methylpiperazino)propane (DUn-MPZ), 3- (N,Ndilinoleylamino)-l ,2-propanediol (DLinAP), 3-(N,N-dioleylamino)-l ,2-propanediol (DOAP), 1 ,2-dilinoleyloxo-3- (2-N,N-dimethylamino)ethoxypropane (DLin-EG-DMA), and 2,2-dilinoleyl-4-dimethylaminomethyl-[l ,3]-dioxoiane (DLin-K-DMA), 2,2-dilinoleyl-4-(2-dimethylaminoethyl)-[l ,3]-dioxoiane (DLki-KC2-DMA); dilinoleyi-methyl-4- dimethylamlnobutyrate (DLin-MC3-DMA); MC3 (US20100324120, which is incorporated herein by reference in its entirety).

[0904] In some embodiments, amino or cationic lipids useful in accordance with the present disclosure have at least one protonatable or deprotonatable group, such that the lipid is positively charged at a pH at or below physiological pH (e.g. pH 7.4), and neutral at a second pH, preferably at or above physiological pH. It will, of course, be understood that the addition or removal of protons as a function of pH is an equilibrium process, and that the reference to a charged or a neutral lipid refers to the nature of the predominant species and does not require that all of lipids have to be present in the charged or neutral form. Lipids having more than one protonatable or deprotonatable group, or which are zwitterionic, are not excluded and may likewise suitable in the context of the present invention. [0905] In some embodiments, a protonatable lipid has a pKa of the protonatable group in the range of about 4 to about 11, e.g., a pKa of about 5 to about 7.

[0906] In some embodiments, a cationic lipid may comprise from about 10 mol % to about 100 mol %, about 20 mol % to about 100 mol %, about 30 mol % to about 100 mol %, about 40 mol % to about 100 mol %, or about 50 mol % to about 100 mol % of total lipid present in a lipid composition utilized in accordance with the present disclosure.

3. Additional lipids or lipid-like materials

[0907] In some embodiments, formulations utilized in accordance with the present disclosure may comprise lipids or lipid-like materials other than cationic or cationically Ionizable lipids or lipid-like materials, i.e., non-cationic lipids or llpid-llke materials (including non-cationically Ionizable lipids or Hpld-llke materials). Collectively, anionic and neutral lipids or lipid-like materials are referred to herein as non-cationic lipids or bpid-like materials. In some embodiments, optimizing a formulation of nucleic acid particles by addition of other hydrophobic moieties, such as cholesterol and lipids, in addition to an ionizable/cationic lipid or lipid-like material may, for example, enhance particle stability and efficacy of nucleic acid delivery.

[0908] In some embodiments, a lipid or lipid-like material may be incorporated which may or may not affect the overall charge of particles. In certain embodiments, such lipid or lipid-like material is a non-cationic lipid or llpid- llke material.

[0909] In some embodiments, a non-cationic lipid may comprise, e.g., one or more anionic lipids and/or neutral lipids. An "anionic lipid" is negatively charged (e.g., at a selected pH).

[0910] A "neutral lipid" exists either in an uncharged or neutral zwitterionic form (e.g., at a selected pH). In some embodiments, a formulation comprises one of the following neutral lipid components: (1) a phospholipid, (2) cholesterol or a derivative thereof; or (3) a mixture of a phospholipid and cholesterol or a derivative thereof. Examples of cholesterol derivatives include, but are not limited to, chdestanol, cholestanone, chdestenone, coprostanol, cholesteryi-2'-hydroxyethyl ether, cholesteryl-4’- hydroxybutyl ether, tocopherol and derivatives thereof, and mixtures thereof.

[0911] Specific exemplary phospholfcids that can be used indude, but are not limited to, phosphatidylcholines, phosphatidylethanolamines, phosphatidylglycerols, phosphatide adds, phosphatidylserines or sphingomyelin. Such phospholipids indude in particular diacylphosphatidylcholines, such as distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dimyristoylphosphatidylcholine (DMPC), dipentadecanoylphosphatidylcholine, dilauroylphosphatidylcholine, dipalmitoylphosphatidylcholine (DPPC), diarachidoylphosphatidykholine (DAPC), dibehenoylphosphatidylcholine (DBPC), ditricosanoylphosphatidylcholine (DTPQ, dilignoceroylphatidylcholine (DLPC), palmltoyloleoyl-phosphatldylchollne (POPC), l,2-di-O-octadeceny1-sn-glycero-3-phosphochoiine (18:0 Diether PC), 1- oteoyl-2<holesterylhemisucdnoyl-sn-glycero-3-phosphocholine (OChemsPC), l-hexadecyl-sn-glycero-3- phosphocholine (C16 Lyso PC) and phosphatidylethanolamines, in particular diacylphosphatidylethanolamines, such as dioleoylphosphatidyiethanolamine (DOPE), distearoyl-phosphatidylethanolamine (DSPE), dipalmitoylphosphatidylethanolamine (DPPE), dimyristoyl-phosphatidylethanolamine (DMPE), dilauroylphosphatidylethanolamine (DLPE), diphytanoyl-phosphatidylethanolamine (DPyPE), and further phosphatidylethanolamine lipids with different hydrophobic chains.

[0912] In certain embodiments, a formulation utilized in accordance with the present disclosure includes DSPC or DSPC and cholesterol. [0913] In certain embodiments, formulations utilized in accordance with the present disclosure include both a cationic lipid and an additional (non-cationic) lipid.

[0914] In some embodiments, formulations herein include a polymer conjugated lipid such as a pegylated lipid. "Pegylated lipids" comprise both a lipid portion and a polyethylene glycol portion. Pegylated lipids are known in the art.

[0915] Without wishing to be bound by theory, the amount of (total) cationic lipid compared to the amount of other lipid(s) in formulation may affect important characteristics, such as charge, particle size, stability, tissue selectivity, and bioactivity of the nucleic add. In some embodiments, the molar ratio of the at least one cationic ipid to the at least one additional lipid is from about 10:0 to about 1:9, about 4:1 to about 1:2, or about 3:1 to about 1:1. [0916] In some embodiments, a non-cationlc lipid, In particular a neutral lipid, (e.g., one or more phospholipids and/or cholesterol) may comprise from about 0 mol % to about 90 mol %, from about 0 mol % to about 80 mol %, from about 0 mol % to about 70 mol %, from about 0 mol % to about 60 mol %, or from about 0 mol % to about 50 mol %, of the total lipid present in a formulation.

4. Lipoptex Partides

[0917] In certain embodiments of the present disclosure, the RNA described herein may be present in RNA lipoplex particles.

[0918] An "RNA lipoplex partide" contains lipid, In particular cationic lipid, and RNA. Electrostatic Interactions between positively charged liposomes and negatively charged RNA results in complexation and spontaneous formation of RNA lipoplex partides. Positively charged liposomes may be generally synthesized using a cationic lipid, such as DOTMA, and additional lipids, such as DOPE. In one embodiment, a RNA lipoplex partide Is a nanopartide. [0919] In certain embodiments, RNA lipoplex particles Include both a cationic lipid and an additional lipid. In an exemplary embodiment, the cationic lipid is DOTMA and the additional lipid Is DOPE.

[0920] In some embodiments, the molar ratio of the at least one cationic lipid to the at least one additional lipid Is from about 10:0 to about 1:9, about 4:1 to about 1:2, or about 3:1 to about 1:1. In specific embodiments, the molar ratio may be about 3:1, about 2.75:1, about 2.5:1, about 2.25:1, about 2:1, about 1.75:1, about 1.5:1, about 1.25:1, or about 1: 1. In an exemplary embodiment, the molar ratio of the at least one cationic lipid to the at least one additional lipid is about 2: 1.

[0921] In some embodiments, RNA lipoplex particles have an average diameter that in one embodiment ranges from about 200 nm to about 1000 nm, from about 200 nm to about 800 nm, from about 250 to about 700 nm, from about 400 to about 600 nm, from about 300 nm to about 500 nm, or from about 350 nm to about 400 nm. In specific embodiments, the RNA lipoplex particles have an average diameter of about 200 nm, about 225 nm, about 250 nm, about 275 nm, about 300 nm, about 325 nm, about 350 nm, about 375 nm, about 400 nm, about 425 nm, about 450 nm, about 475 nm, about 500 nm, about 525 nm, about 550 nm, about 575 nm, about 600 nm, about 625 nm, about 650 nm, about 700 nm, about 725 nm, about 750 nm, about 775 nm, about 800 nm, about 825 nm, about 850 nm, about 875 nm, about 900 nm, about 925 nm, about 950 nm, about 975 nm, or about 1000 nm. In an embodiment, the RNA lipoplex particles have an average diameter that ranges from about 250 nm to about 700 nm. In another embodiment, the RNA lipoplex particles have an average diameter that ranges from about 300 nm to about 500 nm. In an exemplary embodiment, the RNA lipoplex particles have an average diameter of about 400 nm. [0922] RNA lipoplex partides and compositions comprising RNA ipoplex partides described herein are useful for delivery of RNA to a target tissue after parenteral administration, in particular after intravenous administration. The RNA lipoplex partides may be prepared using liposomes that may be obtained by injecting a solution of the lipids in ethanol into water or a suitable aqueous phase. In one embodiment, the aqueous phase has an acidic pH. In one embodiment, the aqueous phase comprises acetic add, e.g., in an amount of about 5 mM. Liposomes may be used for preparing RNA lipoplex partides by mixing the liposomes with RNA. In one embodiment, the Iposomes and RNA lipoplex partides comprise at least one cationic lipid and at least one additional lipid. In one embodiment, the at least one cationic lipid comprises l,2-di-O-octadecenyl-3-trimethylammonium propane (DOTMA) and/or l,2-dioleoyl-3- trimethylammonium-propane (DOTAP). In one embodiment, the at least one additional lipid comprises l,2-di-(9Z- octadecenoyl)-sn-glycero-3-phosphoethanolamlne (DOPE), cholesterol (Choi) and/or l,2-dloleoyl-sn-glycero-3- phosphocholine (DOPC). In one embodiment, the at least one cationic lipid comprises l,2-di-O-octadecenyl-3- trimethylammonium propane (DOTMA) and the at least one additional lipid comprises l,2-di-(9Z-octadecenoyl)-sn- glycero-3-phosphoethanolamine (DOPE). In one embedment, the liposomes and RNA lipoplex particles comprise 1,2- di-O-octadecenyl-3-trimethylammonium propane (DOTMA) and l,2-di-(9Z-octadecenoyl)-sn-glycero-3- phosphoethanolamine (DOPE).

[0923] Spleen targeting RNA lipoplex particles are described In WO 2013/143683, herein incorporated by reference. It has been found that RNA lipoplex particles having a net negative charge may be used to preferentially target spleen tissue or spleen cells such as antigen-presenting cells, In particular dendritic cells. Accordingly, following administration of the RNA lipoplex particles, RNA accumulation and/or RNA expression in the spleen occurs. Thus, RNA lipoplex particles of the disclosure may be used for expressing RNA in the spleen. In an embodiment, after administration of the RNA lipoplex particles, no or essentially no RNA accumulation and/or RNA expression in the lung and/or liver occurs. In one embodiment, after administration of the RNA lipoplex particles, RNA accumulation and/or RNA expression in antigen presenting cells, such as professional antigen presenting cells In the spleen occurs. Thus, RNA lipoplex particles of the disclosure may be used for expressing RNA in such antigen presenting cells. In one embodiment, the antigen presenting cells are dendritic cells and/or macrophages.

5. Lipid Nanopartides (LNPs) [0924] In some embodiments, nucleic add such as RNA described herein Is administered In the form of lipid nanopartides (LNPs). In some embodiments, LNPs may comprise any lipid capable of forming a partide to which the one or more nucleic add molecules are attached, or in which the one or more nucleic acid molecules are encapsulated.

[0925] In some embodiments, an LNP comprises one or more cationic lipids, and one or more stabilizing lipids. Stabilizing lipids indude neutral lipids and pegylated lipids.

[0926] In some embodiments, an LNP comprises a cationic lipid, a neutral lipid, a sterol, a polymer conjugated lipid; and an RNA, encapsulated within or assodated with the lipid nanoparticle.

[0927] In some embodiments, a neutral lipid is selected from the group consisting of DSPC, DPPC, DMPC, DOPC, POPC, DOPE, DOPG, DPPG, POPE, DPPE, DMPE, DSPE, and SM. In some embodiments, the neutral lipid is selected from the group consisting of DSPC, DPPC, DMPC, DOPC, POPC, DOPE and SM. In some embodiments, the neutral lipid is DSPC.

[0928] In some embodiments, a sterol Is cholesterol. [0929] In some embodiments, a polymer conjugated lipid is a pegylated lipid. In some embodiments, a pegylated lipid has the following structure: or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein:

R¹² and R¹³ are each independently a straight or branched, saturated or unsaturated alkyl chain containing from 10 to 30 carbon atoms, wherein the alkyl chain is optionally Interrupted by one or more ester bonds; and w has a mean value ranging from 30 to 60. In some embodiments, R¹² and R¹³ are each independently straight, saturated alkyl chains containing from 12 to 16 carbon atoms. In some embodiments, w has a mean value ranging from 40 to 55. In some embodiments, the average w Is about 45. In some embodiments, R¹² and R¹³ are each Independently a straight, saturated alkyl chain containing about 14 carbon atoms, and w has a mean value of about 45.

[0930] In some embodiments, a pegylated lipid Is DMG-PEG 2000, e.g., having the following structure:

[0931] In some embodiments, a cationic lipid component of LNPs has the structure of Formula (III): or a pharmaceutically acceptable salt, tautomer, prodrug or stereoisomer thereof, wherein: one of L¹ or L² is -0(C=0)-, -<C=0)0-, -C(=0)-, -0-, -5(0),-, -S-S-, -C(=O)S-, SC(=O)-, -NR^aC(=O)-, - C(=O)NR'-, NR^aC(=O)NR^a-, -OC(=O)NR^a- or -NR^aC(=O)O-, and the other of L¹ or L² is -O(C=O)-, -(C=0)0-, -C(=0)- , -0-, -S(O)x-, -S-S-, -C(=O)S-, SC(=O)-, -NR=C(=O)-, -C(=O)NR^a-, NR^aC(=O)NR^a-, -OC(=O)NR^a- or -NR^aC(=O)O- or a direct bond;

G¹ and G² are each independently unsubstituted C₁-C₁₂ alkylene or C₁-C₁₂ alkenylene;

G³ is C₁-C₂₄ alkylene, C₁-C₂₄ alkenylene, C₃-C₈ cycloalkylene, C₃-C₈ cydoalkenylene;

R^a is H or C₁-C₁₂ alkyl;

R¹ and R² are each independently C₅-C₂₄ alkyl or C6-C24 alkenyl;

R³ is H, OR⁵, CN, -C(=O)OR⁴, -OC(=O)R⁴ or -NR⁵C(=O)R⁴;

R⁴ is C₁-C₁₂ alkyl;

R⁵ is H or C₁-C₆ akyl; and x is 0, 1 or 2. [0932] In some of the foregoing embodiments of Formula (III), the lipid has one of the following structures

(IIIA) or (IIIB): wherein:

A is a 3 to 8-membered cycloalkyl or cycloalkylene ring;

R⁶ is, at each occurrence, independently H, OH or Ct-Ca alkyl; and n Is an integer ranging from 1 to 15.

[0933] In some of the foregoing embodiments of Formula (III), the lipid has structure (IIIA), and In other embodiments, the lipid has structure (IIIB).

[0934] In other embodiments of Formula (III), the lipid has one of the following structures (IIIC) or (HID): wherein y and z are each independently integers ranging from 1 to 12.

[0935] In any of the foregoing embodiments of Formula (III), one of L¹ or L² is -0(C=0)-. For example, in some embodiments each of L¹ and L² are -0(C=0)-. In some different embodiments of any of the foregoing, L¹ and L² are each independently -(C=0)0- or -0(C=0)-. For example, in some embodiments each of L¹ and L² is -(C=0)0-

[0936] In some different embodiments of Formula (III), the lipid has one of the following structures (HIE) or

(IIIF):

[0937] In some of the foregoing embodiments of Formula (III), the lipid has one of the following structures

(IIIG), (IIIH), (IIII), or (IID):

[0938] In some of the foregoing embodiments of Formula (III), n is an integer ranging from 2 to 12, for example from 2 to 8 or from 2 to 4. For example, in some embodiments, n is 3, 4, 5 or 6. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5. In some embodiments, n is 6.

[0939] In some other of the foregoing embodiments of Formula (III), y and z are each independently an integer ranging from 2 to 10. For example, in some embodiments, y and z are each independently an integer ranging from 4 to 9 or from 4 to 6.

[0940] In some of the foregoing embodiments of Formula (III), R⁶ is H. In other of the foregoing embodiments, R⁶ is C₁-C₂₄ alkyl. In other embodiments, R⁶ is OH.

[0941] In some embodiments of Formula (III), G³ is unsubstituted. In other embodiments, G3 is substituted.

In various different embodiments, G³ is linear C₁-C₂₄ alkylene or linear C₁-C₂₄ alkenylene.

[0942] In some other foregoing embodiments of Formula (III), R¹ or R², or both, is C₆-C₂₄ alkenyl. For example, in some embodiments, R¹ and R² each, independently have the following structure: wherein:

R⁷¹ and R^7b are, at each occurrence, independently H or C₁-C₁₂ alkyl; and a is an integer from 2 to 12, and wherein R⁷’, R⁷¹¹ and a are each selected such that R¹ and R² each independently comprise from 6 to 20 carbon atoms. For example, in some embodiments a is an integer ranging from 5 to 9 or from 8 to 12.

[0943] In some of the foregoing emtxxiiments of Formula (III), at least one occurrence of R⁷¹ is H. For example, in some embodiments, R⁷¹ is H at each occurrence. In other different embodiments of the foregoing, at least one occurrence of R^7b is C₁-C₈ alkyl. For example, in some embodiments, C₁-C₈ alkyl is methyl, ethyl, n-propyl, iso-propyl, n-butyl, Iso-butyl, tert-butyl, n-hexyl or n-octyl.

[0944] In different embodiments of Formula (III), R¹ or R², or both, has one of the following structures:

[0945] In some of the foregoing embodiments of Formula (III), R³ is OH, CN, -C(=O)OR4, -OC(=O)R⁴ or - NHC(=O)R⁴. In some embodiments, R⁴ is methyl or ethyl.

[0946] In various different embodiments, the cationic lipid of Formula (III) has one of the structures set forth in in Table 26 below.

Table 26: Exemplary Compounds of Formula (III).

[0947] In various different embodiments, a cationic lipid has one of the structures set forth in Table 27 below.

Table 27: Exemplary Cationic Lipid Structures No. Structure

O- .o.

HO.

'N'

E

,O. o

,O,

HO‘ •N"

F

[0948] In some embodiments, an LNP comprises a cationic lipid that is an Ionizable llpid-llke material (lipldold).

In some embodiments, a cationic lipid has the following structure:

[0949] In some embodiments, lipid nanopartides can have an average size (e.g., mean diameter) of about 30 nm to about 150 nm, about 40 nm to about 150 nm, about 50 nm to about 150 nm, about 60 nm to about 130 nm, about 70 nm to about 110 nm, about 70 nm to about 100 nm, about 70 to about 90 nm, or about 70 nm to about 80 nm. In some embodiments, lipid nanopartides in accordance with the present disclosure can have an average size (eg., mean diameter) of about 50 nm to about 100 nm. In some embodiments, lipid nanoparticles may have an average size (e.g., mean diameter) of about 50 nm to about 150 nm. In some embodiments, lipid nanoparticles may have an average size (e.g., mean diameter) of about 60 nm to about 120 nm. In some embodiments, lipid nanopartides in accordance with the present disclosure can have an average size (e.y., mean diameter) of about 30 nm, 35 nm, 40 nm, 45 nm, 50 nm, 55 nm, 60 nm, 65 nm, 70 nm, 75 nm, 80 nm, 85 nm, 90 nm, 95 nm, 100 nm, 105 nm, 110 nm, 115 nm, 120 nm, 125 nm, 130 nm, 135 nm, 140 nm, 145 nm, or 150 nm. The term "average diameter* or "mean diameter" refers to the mean hydrodynamic diameter of particles as measured by dynamic laser light scattering (DLS) with data analysis using the so-called cumulant algorithm, which provides as results the so- called Z-average with the dimension of a length, and the polydispersity Index (PI), which Is dimensionless (Koppel, D., J. Chem. Phys. 57, 1972, pp 4814-4820, ISO 13321, which is herein Incorporated by reference). Here "average diameter," "mean diameter," “diameter," or “size" for particles Is used synonymously with this value of the Z- average.

[0950] In some embodiments, lipid nanopartides described herein may exhibit a polydispersity index less than about 0.5, less than about 0.4, less than about 0.3, or about 0.2 or less. By way of example, lipid nanopartides can exhibit a polydispersity index in a range of about 0.1 to about 0.3 or about 0.2 to about 0.3. The "polydispersity index" is preferably calculated based on dynamic light scattering measurements by the so-called cumulant analysis as mentioned in the definition of the "average diameter." Under certain prerequisites, it can be taken as a measure of the size distribution of an ensemble of ribonudelc add nanopartides (e.g., ribonucleic add nanopartides). [0951] Lipid nanopartides described herein can be characterized by an "N/P ratio/ which is the molar ratio of cationic (nitrogen) groups (the "N" in N/P) in the cationic polymer to the anionic (phosphate) groups (the "P" in N/P) in RNA. It is understood that a cationic group is one that is either in cationic form (e.47_v N⁴), or one that is ionizable to become cationic. Use of a single number in an N/P ratio (api, an N/P ratio of about 5) is intended to refer to that number over 1, e.g., an N/P ratio of about 5 is intended to mean 5:1. In some embodiments, a lipid nanoparticle described herein has an N/P ratio greater than or equal to 5. In some embodiments, a lipid nanoparticle described herein has an N/P ratio that is about 5, 6, 7, 8, 9, or 10. In some embodiments, an N/P ratio for a lipid nanoparticle described herein is from about 10 to about 50. In some embodiments, an N/P ratio for a lipid nanopartide described herein Is from about 10 to about 70. In some embodiments, an N/P ratio for a lipid nanopartide described herein Is from about 10 to about 120.

B. Exemplary Methods of Making Lipid Nanopartides

[0952] Lipids and lipid nanoparticles comprising nucleic adds and their method of preparation are known in the art, induding, e.g., as described in U.S. Patent Nos. 8,569,256, 5,965,542 and U.S. Patent Publication Nos. 2016/0199485, 2016/0009637, 2015/0273068, 2015/0265708, 2015/0203446, 2015/0005363, 2014/0308304, 2014/0200257, 2013/086373, 2013/0338210, 2013/0323269, 2013/0245107, 2013/0195920, 2013/0123338, 2013/0022649, 2013/0017223, 2012/0295832, 2012/0183581, 2012/0172411, 2012/0027803, 2012/0058188, 2011/0311583, 2011/0311582, 2011/0262527, 2011/0216622, 2011/0117125, 2011/0091525, 2011/0076335, 2011/0060032, 2010/0130588, 2007/0042031, 2006/0240093, 2006/0083780, 2006/0008910, 2005/0175682, 2005/017054, 2005/0118253, 2005/0064595, 2004/0142025, 2007/0042031, 1999/009076 and PCT Pub. Nos. WO 99/39741, WO 2018/081480, WO 2017/004143, WO 2017/075531, WO 2015/199952, WO 2014/008334, WO 2013/086373, WO 2013/086322, WO 2013/016058, WO 2013/086373, W02011/141705, and WO 2001/07548, the foil disclosures each of which are herein incorporated by reference In their entirety for the purposes described herein. [0953] For example, in some embodiments, cationic lipids, neutral lipids (e#, DSPC, and/or cholesterol) and polymer-conjugated lipids can be solubilized In ethanol at a pre-determined molar ratio (e.y., ones described herein). In some embodiments, lipid nanopartides (lipid nanopartide) are prepared at a total lipid to polyribonudeotides weight ratio of approximately 10: 1 to 30: 1. In some embodiments, such polyribonudeotides can be diluted to 0.2 mg/mL in acetate buffer.

[0954] In some embodiments, using an ethanol injection technique, a colloidal lipid dispersion comprising polyribonudeotides can be formed as follows: an ethanol solution comprising lipids, such as cationic lipids, neutral lipids, and polymer-conjugated lipids, is injected into an aqueous solution comprising polyribonucleotides (e.gi, ones described herein).

[0955] In some embodiments, lipid and pdyribonudeotide solutions can be mixed at room temperature by pumping each solution at controlled flow rates into a mixing unit, for example, using piston pumps. In some embodiments, the flow rates of a lipid solution and a RNA solution into a mixing unit are maintained at a ratio of 1:3. Upon mixing, nucleic add-lipid particles are formed as the ethanolic ipid solution is diluted with aqueous polyribonudeotides. The lipid solubility is decreased, while cationic lipids bearing a positive charge interact with the negatively charged RNA.

[0956] In some embodiments, a solution comprising RNA-encapsulated lipid nanopartides can be processed by one or more of concentration adjustment, buffer exchange, formulation, and/or filtration.

[0957] In some embodiments, RNA-encapsulated lipid nanoparticles can be processed through filtration. [0958] In some embodiments, particle size and/or internal structure of lipid nanopartides (with or without RNAs) may be monitored by appropriate techniques such as, e.g., small-angle X-ray scattering (SAXS) and/or transmission electron cryomicroscopy (CryoTEM).

V. Pharmaceutical Compositions

[0959] The present disclosure provides compositions, e.g., pharmaceutical compositions comprising one or more polyribonudeotides described herein. Pharmaceutical formulations may additionally comprise a pharmaceutically acceptable excipient, which, as used herein, indudes any and all solvents, dispersion media, diluents, or other liquid vehides, dispersion or suspension aids, surface active agents, isotonic agents, thickening or emulsifying agents, preservatives, solid binders, lubricants and the like, as suited to the particular dosage form desired. Remington's The Science and Practice of Pharmacy, 21st Edition, A. R. Gennaro (Lippincott, WIHIams & Wilkins, Baltimore, MD, 2006; incorporated herein by reference) discloses various exdpients used in formulating pharmaceutical compositions and known techniques for the preparation thereof. Except insofar as any conventional exdpient medium is incompatible with a substance or Its derivatives, such as by produdng any undesirable biological effect or otherwise interacting In a deleterious manner with any other components) of the pharmaceutical composition, its use is contemplated to be within the scope of this disclosure.

[0960] In some embodiments, an excipient is approved for use in humans and for veterinary use. In some embodiments, an excipient Is approved by the United States Food and Drug Administration. In some embodiments, an exdpient is pharmaceutical grade. In some embodiments, an excipient meets the standards of the United States Pharmacopoeia (USP), the European Pharmacopoeia (EP), the British Pharmacopoeia, and/or the International Pharmacopoeia.

[0961] Pharmaceutically acceptable excipients used in the manufacture of pharmaceutical compositions Indude, but are not limited to, Inert diluents, dispersing and/or granulating agents, surface active agents and/or emulsifiers, disintegrating agents, binding agents, preservatives, buffering agents, lubricating agents, and/or oils. Such excipients may optionally be induded In pharmaceutical formulations. Exdpients such as cocoa butter and suppository waxes, coloring agents, coating agents, sweetening, flavoring, and/or perfuming agents can be present in the composition, according to the judgment of the formulator.

[0962] General considerations In the formulation and/or manufacture of pharmaceutical agents may be found, for example, in Remington: The Sdence and Practice of Pharmacy 21st ed., Lippincott Williams & Wilkins, 2005 (incorporated herein by reference).

[0963] In some embodiments, pharmaceutical compositions provided herein may be formulated with one or more pharmaceutically acceptable carriers or diluents as well as any other known adjuvants and exdpients in accordance with conventional techniques such as those disclosed In Remington: The Sdence and Practice of Pharmacy 21st ed., Lippincott Williams & Wilkins, 2005 (incorporated herein by reference).

[0964] Pharmaceutical compositions described herein can be administered by appropriate methods known in the art. As wiN be appredated by a skilled artisan, the route and/or mode of administration may depend on a number of factors, induding, eg., but not limited to stability and/or pharmacokinetics and/or pharmacodynamics of pharmaceutical compositions described herein.

[0965] In some embodiments, pharmaceutical compositions described herein are formulated for parenteral administration, which indudes modes of administration other than enteral and topical administration, usually by injection, and indudes, without limitation, intravenous, intramuscular, intraarterial, intradermal, subcutaneous, subcuticular, or intraarticular injection and infusion. In preferred embodiments, pharmaceutical compositions described herein are formulated for intravenous, intramuscular, or subcutaneous administration. In particularly preferred embodiments, pharmaceutical compositions described herein are formulated for intramuscular administration.

[0966] In some embodiments, pharmaceutical compositions described herein are formulated for intravenous administration. In some embodiments, pharmaceutically acceptable excipients that may be useful for intravenous administration include sterile aqueous solutions or dispersions and sterile powders for preparation of sterile injectable solutions or dispersions.

[0967] Therapeutic compositions typically must be sterile and stable under the conditions of manufacture and storage. The composition can be formulated as a solution, microemulsion, lipid nanopartldes, or other ordered structure suitable to high drug concentration. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof. Proper fluidity can be maintained, for example, by the use of surfactants. In many cases, It wil be preferable to include isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, or sodium chloride In the composition. In some embodiments, prolonged absorption of the injectable compositions can be brought about by including in the composition an agent that delays absorption, for example, monostearate salts and gelatin.

[0968] Sterile injectable solutions can be prepared by incorporating the active compound In the required amount In an appropriate solvent with one or a combination of Ingredients enumerated above, as required, followed by sterilization and/or microfiltration. In some embodiments, pharmaceutical compositions can be prepared as described herein and/or methods known in the art. In some embodiments, a pharmaceutical composition includes ALC-0315; ALC-0159; DSPC; Cholesterol; Sucrose; NaQ; KOI; NajHPO^ KH2PO4; Water for Injection. In some embodiments, normal saline (isotonic 0.9% NaCI) is used as diluent.

[0969] These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents. Prevention of the presence of microorganisms may be ensured both by sterilization procedures, and by the inclusion of various antibacterial and antifungal agents, for example, paraben, chlorobutanol, phenol sorbic acid, and the like. It may also be desirable to include isotonic agents, such as sugars, sodium chloride, and the like into pharmaceutical compositions described herein. In addition, prolonged absorption of the injectable pharmaceutical form may be brought about by the inclusion of agents which delay absorption such as aluminum monostearate and gelatin.

[0970] Formulations of pharmaceutical compositions described herein may be prepared by any method known or hereafter developed In the art of pharmacology. In general, such preparatory methods include the step of bringing active ingredients) into association with a diluent or another excipient and/or one or more other accessory ingredients, and then, if necessary and/or desirable, shaping and/or packaging the product into a desired single- or multi-dose unit

[0971] A pharmaceutical composition in accordance with the present disclosure may be prepared, packaged, and/or sold in bulk, as a single unit dose, and/or as a plurality of single unit doses. As used herein, a “unit dose" is discrete amount of the pharmaceutical composition comprising a predetermined amount of at least one RNA product produced using a system and/or method described herein. [0972] Relative amounts of polyribonucleotides encapsulated in lipid nanopartides, a pharmaceutically acceptable exdpient, and/or any additional ingredients in a pharmaceutical composition can vary, depending upon the subject to be treated, target cells, diseases or disorders, and may also further depend upon the route by which the composition is to be administered.

[0973] In some embodiments, pharmaceutical compositions described herein are formulated into pharmaceutically acceptable dosage forms by conventional methods known to those of skill in the art Actual dosage levels of the active ingredients (e.g., polyribonudeotides encapsulated in lipid nanopartides) in the pharmaceutical compositions described herein may be varied so as to obtain an amount of the active ingredient which is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient. The selected dosage level win depend upon a variety of pharmacokinetic factors Including the activity of the particular compositions of the present disclosure employed, the route of administration, the time of administration, the rate of excretion of the particular compound being employed, the duration of the treatment, other drugs, compounds and/or materials used in combination with the particular compositions employed, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well known in the medical arts.

[0974] A physician having ordinary skill in the art can readily determine and prescribe the effective amount of the pharmaceutical composition required. For example, a physician could start doses of active Ingredients (e.g., polyribonudeotides encapsulated in lipid nanopartides) employed in the pharmaceutical composition at levels lower than that required in order to achieve the desired therapeutic effect and gradually Increase the dosage until the desired effect is achieved.

[0975] In some embodiments, a pharmaceutical composition Is formulated (e.g., but not limited to, for Intravenous, Intramuscular, or subcutaneous administration) to deiver a dose of about 5 mg RNA/kg.

[0976] In some embodiments, a pharmaceutical composition described herein may further comprise one or more additives, for example, In some embodiments that may enhance stability of such a composition under certain conditions. Examples of additives may include but are not limited to salts, buffer substances, preservatives, and carriers. For example, in some embodiments, a pharmaceutical composition may further comprise a cryoprotectant (e.g., sucrose) and/or an aqueous buffered solution, which may In some embodiments Include one or more salts, including, e.g., alkali metal salts or alkaline earth metal salts such as, e.g., sodium salts, potassium salts, and/or calcium salts.

[0977] In some embodiments, a pharmaceutical composition provided herein is a preservative-free, sterile RNA-lipid nanopartide dispersion in an aqueous buffer for intravenous or intramuscular administration.

[0978] Although the descriptions of pharmaceutical compositions provided herein are prindpalty directed to pharmaceutical compositions that are suitable for administration to humans, it will be understood by the skilled artisan that such compositions are generally suitable for administration to animals of all sorts. Modification of pharmaceutical compositions suitable for administration to humans in order to render the compositions suitable for administration to various animals is well understood, and the ordinarily skilled veterinary pharmacologist can design and/or perform such modification with merely ordinary, if any, experimentation.

VI. Patient Populations

[0979] In some aspects, technologies of the present disclosure are used for therapeutic and/or prophylactic purposes. In some embodiments, technologies of the present disclosure are used in the treatment and/or prophylactic of an infection with a malaria parasite. Prophylactic purposes of the present disclosure comprise preexposure prophylaxis and/or post-exposure prophylaxis. In some such embodiments, a malaria parasite is, for example, P. falciparum, P. knowtesi, P. ovate, P. simfovate, P. vivax, P. malariae, P. ovate curtisi, P. ovate waHikeri, and/or P. berghei.

[0980] In some embodiments, technologies of the present disclosure are used in the treatment and/or prophylaxis of a disorder related to such an infection. A disordered related to such an infection comprises, for example, a typical symptom and/or a complication of a malaria infection.

[0981] In some embodiments, provided compositions (e.ff., that are or comprise malarial antigens) may be useful to detect and/or characterize one or more features of an anti-malarial immune response (e.g., by detecting binding to a provided antigen by serum from an Infected subject).

[0982] In some embodiments, provided compositions (e.g., that are or comprise malarial antigens) are useful to raise antibodies to one or more epitopes included therein; such antibodies may themselves be useful, for example for detection or treatment of malarial parasite(s) or infection thereby.

[0983] The present disclosure provides use of encoding nucleic adds (.e.g., DNA or RNA) to produce encoded antigens and/or use of DNA constructs to produce RNA.

[0984] In some embodiments, technologies of the present disclosure are utilized in a non-limlted subject population; in some embodiments, technologies of the present disclosure are utilized In particular subject populations.

[0985] In some embodiments, a subject population comprises an adult population. In some embodiments, an adult population comprises subjects between the ages of about 19 years and about 60 years of age (e.g., about 20, 25, 30, 35, 40, 45, 50, 55, or 60 years of age).

[0986] In some embodiments, a subject population comprises an elderly population. In some embodiments, an elderly population comprises subjects of about 60 years of age, about 70 years of age, or older (e.g., about 65, 70, 75, 80, 85, 90, 95, or 100 years of age).

[0987] In some embodiments, a subject population comprises a pediatric population. In some embodiments, a pediatric population comprises subjects approximately 18 years old or younger. In some such embodiments, a pediatric population comprises subjects between the ages of about 1 year and about 18 years (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 years of age).

[0988] In some embodiments, a subject population comprises a newborn population. In some embodiments, a newborn population comprises subjects about 12 months or younger (e.g., 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1 months or younger). In some embodiments, subject populations to be treated with technologies described herein include infants (e.g., about 12 months or younger) whose mothers did not receive such technologies described herein during pregnancy. In some embodiments, subject populations to be treated with technologies described herein may include pregnant women; in some embodiments, infants whose mothers were treated with disclosed technologies during pregnancy (e.g., who received at least one dose, or alternatively only who received both doses), are not vaccinated during the first weeks, months, or even years (e.g., 1, 2, 3, 4, 5, 6, 7, 8 weeks or more, or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 months or more, or 1, 2, 3, 4, 5 years or more) post-birth. Alternatively or additionally, in some embodiments, infants whose mothers were treated with disclosed technologies during pregnancy (e.g., who received at least one dose, or alternatively only who received both doses), receive reduced treated with disclosed technologies (e.g, lower doses and/or smaller numbers of administrations - e.g, boosters - and/or lower total exposure over a given period of time) after birth, for example during the first weeks, months, or even years (e.g., 1, 2, 3, 4, 5, 6, 7, 8 weeks or more, or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 months or more, or 1, 2, 3, 4, 5 years or more) post-birth or may need reduced vaccination (e.g., lower doses and/or smaller numbers of administrations - e.g., boosters - over a given period of time), In some embodiments, compositions as provided herein are administered to subject populations that do not include pregnant women.

[0989] In some embodiments, a subject population is or comprises children aged 6 weeks to up to 17 months of age.

[0990] In some embodiments, a subject population comprises a population with a high risk of Infection (eg., malaria). In some such embodiments, a population may be deemed to have a high risk of Infection due to a local epidemic or a global pandemic. In some such embodiments, a population may be deemed to have a high risk of infection due to a subject population's geographic area. In some embodiments, a subject population comprises subjects that have been exposed to Infection (eg., malaria).

[0991] In some embodiments, where a subject population is or includes pregnant women, provided technologies offer a particular advantage of interrupting malaria's transmission cycle, including, for example, in some embodiments, by reducing or eliminating transmission from pregnant mothers to their fetuses.

[0992] In some embodiments, a subject population Is or comprises Immunocompromised Individuals. In some embodiments, a subject population does not include immunocompromised individuals.

[0993] In some embodiments, a provided pharmaceutical composition (e.g., Immunogenic composition, e.g., vaccine) may be administered in combination with (l.e., so that subjects) are simultaneously exposed to both) another pharmaceutical composition (e.g., Immunogenic composition, e.g., vaccine) or therapeutic Intervention, e.g., to treat or prevent malaria or another disease, disorder, or condition.

[0994] In some embodiments, a provided pharmaceutical composition (e.g., immunogenic composition, e.g., vaccine) may be administered with a protein vaccine, a DNA vaccine, an RNA vaccine, a cellular vaccine, a conjugate vaccine, etc. In some embodiments, one or more doses of a provided pharmaceutical composition (e.g., immunogenic composition, e.g., vaccine) may be administered together with (e.g., in a single visit) another vaccine or other therapy.

[0995] In some embodiments, a provided pharmaceutical composition (e.g., immunogenic composition, e.g., vaccine) may be administered to subjects who have been exposed, or expect they have been exposed, to malaria. In some embodiments, a provided pharmaceutical composition (e.g., immunogenic composition, e.g., vaccine) may be administered to subjects who do not have symptoms of malarial infection.

VII. Treatment Methods

[0996] In some embodiments, technologies of the present disclosure may be administered to subjects according to a particular dosing regimen. In some embodiments, a dosing regimen may involve a single administration; in some embodiments, a dosing regimen may comprise one or more "booster" administrations after the initial administration. In some embodiments, initial and boost doses are the same amount; in some embodiments they differ. In some embodiments, two or more booster doses are administered. In some embodiments, a plurality of doses are administered at regular intervals. In some embodiments, periods of time between doses become longer. In some embodiments, one or more subsequent doses is administered if a particular clinical (eg., reduction in neutralizing antibody levels) or situational (eg., local development of a new strain) even arises or is detected. [0997] In some embodiments, administered pharmaceutical compositions (e.g., immunogenic compositions, e.g., vaccines) comprising RNA constructs that encode Plasmodium polypeptide constructs are administered in RNA doses of from about 0.1 pg to about 300 pg, about 0.5 pg to about 200 pg, or about 1 pg to about 100 pg, such as about 1 pg, about 3 pg, about 10 pg, about 30 pg, about 50 pg, or about 100 pg. In some embodiments, an saRNA construct is administered at a lower dose (e.p., 2, 4, 5, 10 fold or more lower) than a modRNA or uRNA construct [0998] In some embodiments, a first booster dose is administered within a about six months of the initial dose, and preferably within about 5, 4, 3, 2, or 1 months. In some embodiments, a first booster dose is administered in a time period that begins about 1, 2, 3, or 4 weeks after the first dose, and ends about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 weeks of the first dose (e.g., between about 1 and about 12 weeks after the first dose, or between about 2 or 3 weeks and about 5 and 6 weeks after the first dose, or about 3 weeks or about 4 weeks after the first dose). [0999] In some embodiments, a plurality of booster doses (e.g., 2, 3, or 4) doses are administered within 6 months of the first dose, or within 12 months of the first dose.

[1OOO] In some embodiments, 3 doses or fewer are required to achieve effective vaccination (e.g., greater than 60%, and in some embodiments greater than about 70%, about 75%, about 80%, about 85%, about 90% or more) reduction in risk of infection, or of serious disease. In some embodiments, not more than two doses are required. In some embodiments, a single dose is sufficient. In some embodiments, an RNA dose is about 60 pg or lower, 50 pg or lower, 40 pg or lower, 30 pg or lower, 20 pg or lower, 10 pg or lower, 5 pg or lower, 2.5 pg or lower, or 1 pg or lower. In some embodiments, an RNA dose is about 0.25 pg, at least 0.5 pg, at least 1 pg, at least 2 pg, at least 3 pg, at least 4 pg, at least 5 pg, at least 10 pg, at least 20 pg, at least 30 pg, or at least 40 pg. In some embodiments, an RNA dose is about 0.25 pg to 60 pg, 0.5 pg to 55 pg, 1 pg to 50 pg, 5 pg to 40 pg, or 10 pg to 30 pg may be administered per dose In some embodiments, an RNA dose Is about 30 pg. In some embodiments, at least two such doses are administered. For example, a second dose may be administered about 21 days following administration of the first dose. In some embodiments, a first booster dose Is administered about one month after an Initial dose. In some such embodiments, at least one further booster is administered at one-month Intervals). In some embodiments, after 2 or 3 boosters, a longer interval Is Introduced and no further booster is administered for at least 6, 9, 12, 18, 24, or more months. In some embodiments, a single further booster is administered after about 18 months. In some embodiments, no further booster Is required unless, for example, a material change in clinical or environmental situation is observed. vin. Methods of Manufacture

[1001] Individual polyribonucleotides can be produced by methods known in the art For example, in some embodiments, polyribonucleotides can be produced by in vitro transcription, for example, using a DNA template. A plasmid DNA used as a template for in vitro transcription to generate a polyribonucleotide described herein Is also within the scope of the present disclosure.

[1002] A DNA template is used for in vitro RNA synthesis in the presence of an appropriate RNA polymerase (e.g., a recombinant RNA-polymerase such as a T7 RNA-polymerase) with ribonucleotide triphosphates (e.g., ATP, CTP, GTP, DTP). In some embodiments, polyribonucleotides (e.g., ones described herein) can be synthesized in the presence of modified ribonucleotide triphosphates. By way of example only, in some embodiments, pseudouridine (Ψ ), Nl-methyl-pseudouridine (mlqr), or 5-methyl-uridine (m5U) can be used to replace uridine triphosphate (UTP). In some embodiments, pseudouridine (Ψ) can be used to replace uridine triphosphate (UTP). In some embodiments,

AAAAAAAAAAAAAAAAAAAAAAAA (SEQ ID NO: 112). In some embodiments, a poly(A) tail comprises a plurality of A embodiments may be or comprise magnetic bead-based chromatography. In some embodiments, polyribonucleotides may be purified using hydrophobic interaction chromatography (HIC) and/or diafiltration. In some embodiments, polyribonucleotides may be purified using HIC followed by cfiafiltration.

[1009] In some embodiments, dsRNA may be obtained as side product during in vitro transcription. In some such embodiments, a second purification step may be performed to remove dsRNA contamination. For example, in some embodiments, cellulose materials (e.g, microcrystalline celldose) may be used to remove dsRNA contamination, for examples in some embodiments in a chromatographic format. In some embodiments, cellulose materials (e.y., microcrystalline cellulose) can be pretreated to inactivate potential RNase contamination, for example in some embodiments by autoclaving followed by incubation with aqueous basic solution, e.g., NaOH. In some embodiments, cellulose materials may be used to purify polyribonucleotides according to methods described In WO 2017/182524, the entire content of which is incorporated herein by reference.

[1010] In some embodiments, a batch of polyribonucleotides may be further processed by one or more steps of filtration and/or concentration. For example, in some embodiments, pdyribonucleotide(s), for example, after removal of dsRNA contamination, may be further subject to diafiltration (6.47., in some embodiments by tangential flow filtration), for example, to adjust the concentration of polyribonucleotides to a desirable RNA concentration and/or to exchange buffer to a drug substance buffer.

[1011] In some embodiments, polyribonucleotides may be processed through 0.2 pm filtration before they are filled into appropriate containers.

[1012] In some embodiments, polyribonucleotides and compositions thereof may be manufactured in accordance with a process as descrfoed herein, or as otherwise known in the art.

[1013] In some embodiments, polyribonucleotides and compositions thereof may be manufactured at a large scale. For example, in some embodiments, a batch of polyribonucleotides can be manufactured at a scale of greater than 1 g, greater than 2 g, greater than 3 g, greater than 4 g, greater than 5 g, greater than 6 g, greater than 7 g, greater than 8 g, greater than 9 g, greater than 10 g, greater than 15 g, greater than 20 g, or higher.

[1014] In some embodiments, RNA quality control may be performed and/or monitored at any time during production process of polyribonucleotides and/or compositions comprising the same. For example, in some embodiments, RNA quality control parameters, Including one or more of RNA identity (e.ry., sequence, length, and/or RNA natives), RNA integrity, RNA concentration, residual DNA template, and residual dsRNA, may be assessed and/or monitored after each or certain steps of a polyribonucleotide manufacturing process, e.g., after in vitro transcription, and/or each purification step.

[1015] In some embodiments, the stability of polyribonucleotides (e.47., produced by in vitro transcription) and/or compositions comprising polyribonucleotides can be assessed under various test storage conditions, for example, at room temperatures vs. fridge or sub-zero temperatures over a period of time (e.g., at least 3 months, at least 6 months, at least 9 months, at least 12 months, or longer). In some embodiments, polyribonucleotides (e.g., ones described herein) and/or compositions thereof may be stored stable at a fridge temperatire (e.g., about 4°C to about 10°C) for at least 1 month or longer Including, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, or at least 12 months or longer. In some embodiments, polyribonucleotides (e.g., ones described herein) and/or compositions thereof may be stored stable at a sub-zero temperature (eg., -20°C or below) for at least 1 month or longer Including, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, or at least 12 months or longer. In some embodiments, polyribonucleotides (eg., ones described herein) and/or compositions thereof may be stored stable at room temperature (eg., at about 25°C) for at least 1 month or longer.

[1016] In some embodiments, one or more assessments may be utilized during manufacture, or other preparation or use of polyribonucleotides (eg., as a release test).

[1017] In some embodiments, one or more quality control parameters may be assessed to determine whether polyribonucleotides described herein meet or exceed acceptance criteria (eg., for subsequent formulation and/or release for distribution). In some embodiments, such quality control parameters may include, but are not limited to RNA integrity, RNA concentration, residual DNA template and/or residual dsRNA. Certain methods for assessing RNA quality are known in the art; for example, one of skill In the art will recognize that In some embodiments, one or more analytical tests can be used for RNA quality assessment. Examples of such certain analytical tests may include but are not limited to gel electrophoresis, UV absorption, and/or PCR assay.

[1018] In some embodiments, a batch of polyribonucleotides may be assessed for one or more features as described herein to determine next action step(s). for example, a batch of polyribonucleotides can be designated for one or more further steps of manufacturing and/or formulation and/or distribution if RNA quality assessment indicates that such a batch of polyribonucleotides meet or exceed the relevant acceptance criteria. Otherwise, an alternative action can be taken (eg., discarding the batch) If such a batch of polyribonucleotides does not meet or exceed the acceptance criteria.

[1019] In some embodiments, a batch of polyribonucleotides that satisfy assessment results can be utilized for one or more further steps of manufacturing and/or formulation and/or distribution.

DC. DNA Constructs

[1020] Among other things, the present disclosure provides DNA constructs, for example that may encode one or more antibody agents as described herein, or components thereof. In some embodiments, DNA constructs provided by and/or utilized in accordance with the present disclosure are comprised In a vector.

[1021] Non-limiting examples of a vector Include plasmid vectors, cosmid vectors, phage vectors such as lambda phage, viral vectors such as retroviral, adenoviral or baculovlral vectors, or artificial chromosome vectors such as bacterial artificial chromosomes (BAC), yeast artificial chromosomes (YAC), or Pl artificial chromosomes (PAC). In some embodiments, a vector is an expression vector. In some embodiments, a vector is a cloning vector. In general, a vector is a nucleic acid construct that can receive or otherwise become linked to a nucleic acid element of interest (eg., a construct that is or encodes a payload, or that imparts a particular functionality, etc.).

[1022] Expression vectors, which may be plasmid or viral or other vectors, typically include an expressible sequence of interest (e.g., a coding sequence) that is functionally linked with one or more control elements (e.g., promoters, enhancers, transcription terminators, etc.). Typically, such control elements are selected for expression in a system of interest. In some embodiments, a system is ex vivo (e.g., an in vitro transcription system); in some embodiments, a system is #? vivo (e.g., a bacterial, yeast, plant, insect, fish, vertebrate, mammalian cell or tissue, etc.).

[1023] Cloning vectors are generally used to modify, engineer, and/or duplicate (eg., by replication in vivo, for example in a simple system such as bacteria or yeast, or i? vitro, such as by amplification such as polymerase chain reaction or other amplification process). In some embodiments, a cloning vector may lack expression signals. [1024] In many embodiments, a vector may indude replication elements such as primer binding site(s) and/or origin(s) of replication. In many embodiments, a vector may indude insertion or modification sites such as restriction endonuclease recognition sites and/or guide RNA binding sites, eta

[1025] In some embodiments, a vector is a viral vector (eg., an AAV vector). In some embodiments, a vector is a non-viral vector. In some embodiments, a vector is a plasmid.

[1026] Those skilled in the art are aware of a variety of technologies useful for the production of recombinant polynucleotides (eg., DNA or RNA) as described herein. For example, restriction digestion, reverse transcription, amplification (eg., by polymerase chain reaction), Gibson assembly, etc., are well established and useful tools and technologies. Alternatively or additionally, certain nudeic acids may be prepared or assembled by chemical and/or enzymatic synthesis. In some embodiments, a combination of known methods is utilized to prepare a recombinant polynucleotide.

[1027] In some embodiments, polynudeotide(s) of the present disclosure are induded in a DNA construct

(eg., a vector) amenable to transcription and/or translation.

[1028] In some embodiments, an expression vector comprises a polynudeotide that encodes proteins and/or polypeptides of the present disdosure operatively linked to a sequence or sequences that control expression (eg., promoters, start signals, stop signals, polyadenylation signals, activators, repressors, eft:). In some embodiments, a sequence or sequences that control expression are selected to achieve a desired level of expression. In some embodiments, more than one sequence that controls expression (eg., promoters) are utilized. In some embodiments, more than one sequence that controls expression (eg., promoters) are utilized to achieve a desired level of expression of a plurality of polynucleotides that encode a plurality proteins and/or polypeptides. In some embodiments, a plurality of recombinant proteins and/or polypeptides are expressed from the same vector (eg., a bi-cistronic vector, a tri-dstronic vector, multi-dstronlc). In some embodiments, a plurality of polypeptides are expressed, each of which Is expressed from a separate vector.

[1029] In some embodiments, an expression vector comprising a polynudeotide of the present disclosure Is used to produce a RNA and/or protein and/or polypeptide in a host cell. In some embodiments, a host cell may be in vitro (eg., a cell line) - for example a cell or cell line (eg., Human Embryonic Kidney (HEK cells), Chinese Hamster Ovary cells, etc) suitable for producing polynucleotides of the present disclosure and proteins and/or polypeptides encoded by said polynudeotides.

[1030] In some embodiments, an expression vector is an RNA expression vector. In some embodiments, an RNA expression vector comprises a polynucleotide template used to produce a RNA in cell-free enzymatic mix. In some embodiments, an RNA expression vector comprising a polynucleotide template is enzymatically linearized prior to In vitro transcription. In some embodiments, a polynucleotide template Is generated through PCR as a linear polynudeotide template. In some embodiments, a linearized polynudeotide is mixed with enzymes suitable for RNA synthesis, RNA capping and/or purification. In some embodiments, the resulting RNA is suitable for producing proteins encoded by the RNA.

[1031] A variety of methods are known in the art to introduce an expression vector into host cells. In some embodiments, a vector may be introduced into host cells using transfection. In some embodiments, transfection is completed, for example, using calcium phosphate transfection, lipofection, or polyethylenimine- mediated transfection. In some embodiments, a vector may be introduced into a host cell using transduction. [1032] In some embodiments, transformed host cells are cultured following introduction of a vector into a host cel to allow for expression of said recombinant polynucleotides. In some embodiments, a transformed host cells are cultured for at least 12 hours, 16 hours, 20 hours, 24 hours, 28 hours, 32 hours, 36 hours 40 hours, 44 hours, 48 hours, 52 hours, 56 hours, 60 hours, 64 hours, 68 hours, 72 hours or longer. Transformed host cells are cultured in growth conditions (e.g., temperature, carbon-dioxide levels, growth medium) in accordance with the requirements of a host cell selected. A skilled artisan would recognize culture conditions for host cells selected are well known in the art.

EXEMPLARY NUMBERED EMBODIMENTS

[1033] Embodiment 1: A polyribonucleotide encoding a polypeptide, wherein the polypeptide comprises one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof.

[1034] Embodiment 2: The polyribonucleotide of embodiment 1, wherein the one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof comprise:

(i) one or more Plasmodium reticulocyte-binding protein homolog 5 (Rh5) polypeptides or antigenic portions thereof;

(ii) one or more Plasmodium Cysteine-Rich Protective Antigen (CyRPA) polypeptides or antigenic portions thereof;

(iii) one or more Plasmodium Rh5-interacting Protein (Ripr) polypeptides or antigenic portions thereof;

(iv) one or more Plasmodium P113 polypeptides or antigenic portions thereof;

(v) one or more Plasmodium thrombospondin-related apical merozoite protein (TRAMP) polypeptides or antigenic portions thereof; or

(vi) one or more Plasmodium cysteine-rich small secreted protein (CSS) polypeptides or antigenic portions thereof.

[1035] Embodiment 3: The polyribonucleotide of embodiment 1 or 2, wherein the one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof comprises one or more Plasmodium reticulocytebinding protein homolog 5 (Rh5) polypeptides or antigenic portions thereof.

[1036] Embodiment 4: The polyribonucleotide of any one of embodiments 1-3, wherein the one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof comprise one or more antigenic portions of Plasmodium Rh5.

[1037] Embodiment 5: The polyribonucleotide of any one of embodiments 2-4, wherein the one or more Plasmodium Rh5 antigenic portions comprise one or more ordered domains of Rh5.

[1038] Embodiment 6: The polyribonucleotide of any one of embodiments 2-5, wherein the one or more Plasmodium Rh5 antigenic portions comprise two ordered domains of Rh5.

[1039] Embodiment 7: The polyribonucleotide of embodiment 6, wherein the two ordered domains of Rh5 are directly adjacent to one another.

[1040] Embodiment 8: The polyribonucleotide of any one of embodiments 2-7, wherein the one or more Plasmodium Rh5 antigenic portions comprise a cysteine at position 203, 329, or both, as numbered according to SEQ ID NO: 1.

[1041] Embodlment 9: The polyribonucleotide of embodiment 8, wherein the one or more Plasmodium Rh5 antigenic portions comprise: (i) an amino add sequence according to SEQ ID NO: 118, 119, 124, or 126,

(ii) an amino acid sequence according to SEQ ID NO: 120 or 128, or

(iii) a combination thereof.

[1042] Embodiment 10: The polyribonucleotide of any one of embodiments 2-7, wherein the one or more

Plasmodium Rh5 antigenic portions comprise a tyrosine at position 203, 329, or both, as numbered according to SEQ

ID NO: 1.

[1043] Embodiment 11: The polyribonucleotide of embodiment 10, wherein the one or more Plasmodium

Rh5 antigenic portions comprise:

(i) an amino acid sequence according to SEQ ID NO: 121, 122, 125, or 127,

(II) an amino acid sequence according to SEQ ID NO: 123 or 129, or

(Hi) a combination thereof.

[1044] Embodiment 12: The polyribonucleotide of any one of embodiments 2-8, 10, and 11, wherein one of the Plasmodium Rh5 antigenic portions comprise a tyrosine at position 203, as numbered according to SEQ ID NO: 1.

[1045] Embodiment 13: The polyribonucleotide of any one of embodiments 2-8 and 10-12, wherein one of the Plasmodium Rh5 antigenic portions comprise a tyrosine at position 329, as numbered according to SEQ ID NO: 1.

[1046] Embodiment 14: The polyribonucleotide of any one of embodiments 2-13, wherein the one or more Plasmodium Rh5 antigenic portions comprise one, two, three, or four N-llnked glycosylation sites.

[1047] Embodiment 15: The polyribonucleotide of any one of embodiments 2-14, wherein the one or more Plasmodium Rh5 antigenic portions comprise two N-llnked glycosylation sites.

[1048] Embodiment 16: The polyribonucleotide of any one of embodiments 2-15, wherein the one or more Plasmodium Rh5 antigenic portions comprise an amino add substitution at one or more N-llnked glycosylation sites, wherein the amino add substitution prevents glycosylation.

[1048] Embodiment 17: The polyribonucleotide of any one of embodiments 2-16, wherein the one or more Plasmodium Rh5 antigenic portions comprise a substitution of NX[T/S] to QX[T/S] and/or a substitution of NX[T/S] to NXA.

[1050] Embodiment 18: The polyribonucleotide of any one of embodiments 2-17, wherein the one or more

Plasmodium Rh5 antlgeric portions comprise:

(i) an amino add sequence according to SEQ ID NO: 124, 125, 126, or 127,

(ii) an amino acid sequence according to SEQ ID NO: 128 or 129, or

(iii) a combination thereof.

[1051] Embodiment 19: The polyribonucleotide of any one of embodiments 2-18, wherein the one or more

Plasmodium Rh5 antigenic portions comprise an amino add substitution that prevents glycosylation at position 214, as numbered according to SEQ ID NO: 1.

[1052] Embodiment 20: The polyribonucleotide of any one of embodiments 2-19, wherein the one or more Plasmodium Rh5 antigenic portions comprise an amino add substitution that prevents glycosylation at position 297, as numbered accorcfing to SEQ ID NO: 1.

[1053] Embodiment 21: The polyribonucleotide of any one of embodiments 2-20, wherein the one or more Plasmodium Rh5 antigenic portions comprise an amino add substitution at all of the N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation. [1054] Embodiment 22: The polyribonudeotide of any one of embodiments 2-21, wherein the one or more Plasmodium Rh5 antigenic portions comprise a PMX deavage site.

[1055] Embodiment 23: The polyribonucleotide of any one of embodiments 2-22, wherein the one or more

Plasmodium Rh5 antigenic portions comprise an amino add sequence according to SEQ ID NO: 118, 121, 124, or 125.

[1056] Embodiment 24: The polyribonucleotide of any one of embodiments 2-21, wherein the one or more Plasmodium Rh5 antigenic portions do not comprise a PMX cleavage site.

[1057] Embodiment 25: The polyribonucleotide of any one of embodiments 2-21 and 24, wherein the one or more Plasmodium Rh5 antigenic portions comprise an amino acid sequence according to SEQ ID NO: 119, 122, 126, or 127.

[1058] Embodiment 26: The polyribonucleotide of embodiment 5, wherein the one or more Plasmodium Rh5 antigenic portions comprise:

(i) an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 130,

(ii) an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 118, 119, 121, 122, 124, 125, 126, or 127,

(Hi) an amino add sequence with at least 85% sequence Identity to an amino acid sequence according to SEQ ID NO: 131,

(Iv) an amino add sequence with at least 85% sequence Identity to an amino acid sequence according to SEQ ID NO: 120, 123, 128, or 129, or (v) a combhation thereof.

[1059] Embodiment 27: The polyribonucleotide of any one of embodiments 1-26, wherein the one or more

Plasmodium Rh5 antigenic portions comprise:

(I) an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 118, 119, 121, 122, 124, 125, 126, or 127,

(II) an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 120, 123, 128, or 129, or

(Hi) a combination thereof.

[1060] Embodiment 28: The polyribonucleotide of any one of embodiments 1-27, wherein the one or more

Plasmodium Rh5 antigenic portions comprise:

(i) an amino add sequence according to SEQ ID NO: 121 or 122, and

(il) an amino add sequence according to SEQ ID NO: 120.

[1061] Embodiment 29: The polyribonucleotide of any one of embodiments 1-27, wherein the one or more

Plasmodium Rh5 antigenic portions comprise:

(i) an amino acid sequence according to SEQ ID NO: 125 or 127, and

(ii) an amino acid sequence according to SEQ ID NO: 120 or 128.

[1062] Embodiment 30: The polyribonucleotide of any one of embodiments 1-4, wherein the polypeptide comprises an amino add sequence with at least 85% sequence identity to an amino add sequence of positions 26- 363 of SEQ ID NO: 94, 95 or 99. [1063] Embodiment 31: The polyribonucleotide of any one of embodiments 1-4, wherein the polypeptide comprises an amino add sequence with at least 85% sequence identity to an amino acid sequence of positions 26- 526 of SEQ ID NO: 89.

[1064] Embodiment 32: The polyribonucleotide of any one of embodiments 1-4, wherein the polypeptide comprises an amino add sequence of positions 26-363 of SEQ ID NO: 94.

[1065] Embodiment 33: The polyribonucleotide of any one of embodiments 1-4, wherein the polypeptide comprises an amino add sequence of positions 26-363 of SEQ ID NO: 95.

[1066] Embodiment 34: The polyribonucleotide of any one of embodiments 1-4, wherein the polypeptide comprises an amino add sequence of positions 26-363 of SEQ ID NO: 99.

[1067] Embodiment 35: The polyribonucleotide of any one of embodiments 1-4, wherein the polypeptide comprises an amino add sequence of positions 26-526 of SEQ ID NO: 89.

[1068] Embodiment 36: The polyribonucleotide of any one of embodiments 1-4, wherein the one or more Plasmodium Rh5 antigenic portions do not comprise a disordered domain of Rh5.

[1069] Embodiment 37: The polyribonucleotide of any one of embodiments 22-29, wherein the PMX deavage site comprises or consists of an amino acid sequence of NFLQ.

[1070] Embodiment 38: The polyribonucleotide of embodiment 1 or 2, wherein the one or more Plasmodium Rh5 Invasion complex polypeptides or antigenic portions thereof comprise a PlasmodlumCy^Pk polypeptide or antigenic portion thereof.

[1071] Embodiment 39: The polyribonucleotide of embodiment 38, wherein the one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof comprise an antigenic portion of Plasmodium CyRPA.

[1072] Embodiment 40: The polyribonucleotide of embodiment 39, wherein the Plasmodium CyRPA antigenic portion comprises of an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 133.

[1073] Embodiment 41: The polyribonudeotide of embodiment 39 or 40, wherein the Plasmodium CyRPA antigenic portion comprises or consists of an amino add sequence of SEQ ID NO: 133.

[1074] Embodiment 42: The polyribonudeotide of any one of embodiments 39-41, wherein the Plasmodium CyRPA antigenic portion comprises of an amino add sequence with at least 85% sequence Identity to an amino acid sequence according to SEQ ID NO: 169.

[1075] Embodiment 43: The polyribonucleotide of any one of embodiments 39-42, wherein the Plasmodium CyRPA antigenic portion comprises or consists of an amino acid sequence of SEQ ID NO: 169.

[1076] Embodiment 44: The polyribonucleotide of any one of embodiments 39-43, wherein the Plasmodium CyRPA antigenic portion comprises one, two, or three N-linked glycosylation sites.

[1077] Embodiment 45: The polyribonucleotide of embodiment 44, wherein the Plasmodium CyRPA antigenic portion comprises an amino add substitution at one or more N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation.

[1078] Embodiment 46: The polyribonudeotide of any one of embodiments 39, 44, and 45, wherein the Plasmodium CyRPA antigenic portion comprises a substitution of NX[T/S] to QX[T/S],

[1079] Embodiment 47: The polyribonucleotide of any one of embodiments 39 and 44-46, wherein the Plasmodium CyRPA antigenic portion comprises a substitution of NX(T/S] to NXA. [1080] Embodiment 48: The polyribonucleotide of any one of embodiments 44-47, wherein the Plasmodium CyRPA antigenic portion comprises an amino add substitution at all of the N-linked glycosylation sites, wherein the amino acid substitution prevents glycosylation.

[1081] Embodiment 49: The polyribonucleotide of any one of embodiments 39-47, wherein the Plasmodium CyRPA antigenic portion comprises an asparagine at position 145, position 322, position 337, or a combination thereof, as numbered according to SEQ ID NO: 3.

[1082] Embodiment 50: The polyribonucleotide of any one of embodiments 39-48, wherein the Plasmodium CyRPA antigenic portion comprises a glutamine at position 145, position 322, position 337, or a combination thereof, as numbered according to SEQ ID NO: 3.

[1083] Embodiment 51: The polyribonucleotide of any one of embodiments 39-48 or 50, wherein the Plasmodium CyRPA antigenic portion comprises a glutamine at position 145, position 322, position 337, as numbered according to SEQ ID NO: 3.

[1084] Embodiment 52: The polyribonucleotide of any one of embodiments 45-48, 50 and 51, wherein the Plasmodium CyRPA antigenic portion comprises or consists of an amino add sequence of SEQ ID NO: 134.

[1085] Embodiment 53: The polyribonucleotide of any one of embodiments 45-48, 50 and 51, wherein the Plasmodium CyRPA antigenic portion comprises or consists of an amino acid sequence of SEQ ID NO: 170.

[1086] Embodiment 54: The polyribonucleotide of embodiment 1 or 2, wherein the one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof comprise a Plasmodium P113 polypeptide or antigenic portions thereof.

[1087] Embodiment 55: The polyribonucleotide of embodiment 54, wherein the one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof comprise an antigenic portion of Plasmodium Pl 13.

[1088] Embodiment 56: The polyribonucleotide of embodiment 55, wherein the Piasmocflum P113 antigenic portion comprises an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 135.

[1089] Embodiment 57: The polyribonudeotide of embodiment 55 or 56, wherein the Plasmodium P113 antigenic portion comprises or consists of an amino add sequence of SEQ ID NO: 135.

[1090] Embodiment 58: The polyribonucleotide of embodiment 55, wherein the Piasmocfium P113 antigenic portion comprise one, two, three, four, five, six, seven, or eight N-linked glycosylation sites.

[1091] Embodiment 59: The polyribonucleotide of embodiment 55 or 58, wherein the Plasmodium P113 antigenic portion comprises an amino acid substitution at one or more N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation.

[1092] Embodiment 60: The polyribonucleotide of any one of embodiments 55, 58 and 59, wherein the Plasmodium Pl 13 antigenic portion comprises a substitution of NX[T/S] to QX[T/S].

[1093] Embodiment 61: The polyribonudeotide of any one of embodiments 55 and 58-60, wherein the Plasmodium Pl 13 antigenic portion comprises a substitution of NX[T/S] to NXA.

[1094] Embodiment 62: The polyribonudeotide of any one of embodiments 55 and 58-61, wherein the Plasmodium P113 antigenic portion comprises an amino acid substitution at all of the N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation. [1095] Embodiment 63: The polyribonucleotide of any one of embodiments 55-61, wherein the Plasmodium P113 antigenic portion comprises an asparagine at position 207, position 268, position 317, position 360, position 661, position 697, position 779, position 876, or a combination thereof, as numbered according to SEQ ID NO: 6. [1096] Embodiment 64: The polyribonucleotide of any one of embodiments 55, 58-61, and 63, wherein the Plasmodium P113 antigenic portion comprises a glutamine at position 207, position 268, position 317, position 360, position 661, position 697, position 779, position 876, or a combination thereof, as numbered according to SEQ ID NO: 6.

[1097] Embodiment 65: The polyribonucleotide of any one of embodiments 55, 58-61, 63, and 64, wherein the Plasmodium Pl 13 antigenic portion comprises a glutamine position 207, position 268, position 317, position 360, position 661, position 697, position 779, and position 876, as numbered according to SEQ ID NO: 6.

[1098] Embodiment 66: The polyribonucleotide of any one of embodiments 55, 58-61, and 63-65, wherein the Plasmodium Pl 13 antigenic portion thereof comprises or consists of an amino add sequence of SEQ ID NO: 136.

[1099] Embodiment 67: The polyribonucleotide of embodiment 1 or 2, wherein the one or more Plasmodium Rh5 Invasion complex polypeptides or antigenic portions thereof comprise a Plasmodium Ripr polypeptide or antigenic portion thereof.

[1100] Embodiment 68: The polyribonucleotide of embodiment 67, wherein the one or more Plasmodium Rh5 Invasion complex polypeptides or antigenic portions thereof comprise an antigenic portion of Plasmodium Ripr.

[1101] Embodiment 69: The polyribonucleotide of embodiment 68, wherein the Plasmodium Ripr antigenic portion comprises or consists of an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 178.

[1102] Embodiment 70: The polyribonucleotide of embodiment 68 or 69, wherein the Plasmodium Ripr antigenic portion comprises or consists of an amino add sequence according to SEQ ID NO: 178.

[1103] Embodiment 71: The polyribonucleotide of embodiment 68, wherein the Plasmodium Ripr antigenic portion comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 177.

[1104] Embodiment 72: The polyribonucleotide of embodiment 68 or 71, wherein the Plasmodium Ripr antigenic portion comprises or consists of an amino acid sequence according to SEQ ID NO: 177.

[1105] Embodiment 73: The polyribonucleotide of embodiment 68, wherein the Plasmodium Ripr antigenic portion comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 176.

[1106] Embodiment 74: The polyribonucleotide of embodiment 68 or 73, wherein the Plasmodium Ripr antigenic portion comprises or consists of an amino acid sequence according to SEQ ID NO: 176.

[1107] Embodiment 75: The polyribonucleotide of embodiment 68, wherein the Plasmodium Ripr antigenic portion comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 175.

[1108] Embodiment 76: The polyribonucleotide of embodiment 68 or 75, wherein the Plasmodium Ripr antigenic portion comprises or consists of an amino acid sequence according to SEQ ID NO: 175.

[1109] Embodiment 77: The polyribonucleotide of embodiment 68, wherein the Plasmodium Ripr antigenic portion comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 174. [1110] Embodiment 78: The polyribonucleotide of embodiment 68 or 77, wherein the Plasmodium Ripr antigenic portion comprises or consists of an amino acid sequence according to SEQ ID NO: 174.

[1111] Embodiment 79: The polyribonucleotide of embodiment 68, wherein the Plasmodium Ripr antigenic portion comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 190.

[1112] Embodiment 80: The polyribonucleotide of embodiment 68 or 79, wherein the Plasmodium Ripr antigenic portion comprises or consists of an amino acid sequence according to SEQ ID NO: 190.

[1113] Embodiment 81: The polyribonucleotide of embodiment 68, wherein the Plasmodium Ripr antigenic portion comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 171.

[1114] Embodiment 82: The polyribonucleotide of embodiment 68 or 81, wherein the Plasmodium Ripr antigenic portion comprises or consists of an amino add sequence according to SEQ ID NO: 171.

[1115] Embodiment 83: The polyribonucleotide of any one of embodiments 68-82, wherein the Plasmodium Ripr antigenic portion comprises one, two, three, four, five, six, seven, eight, nine, ten, eleven or twelve N-linked glycosylation sites.

[1116] Embodiment 84: The polyribonucleotide of embodiment 83, wherein the Piasmocfium Ripr antigenic portion comprises an amino add substitution at one or more N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation.

[1117] Embodiment 85: The polyribonucleotide of any one of embodiments 68-84, wherein the Plasmodium

Ripr antigenic portion comprises a substitution of NX[T/S] to QX[T/S],

[1118] Embodiment 86: The polyribonucleotide of any one of embodiments 83-85, wherein the Plasmodium Ripr antigenic portion comprises an amino acid substitution at all of the N-linked glycosylation sites, wherein the amino acid substitution prevents glycosylation.

[1119] Embodiment 87: The polyribonucleotide of any one of embodiments 68-86, wherein the Plasmodium Ripr antigenic portion comprises an asparagine at position 646, position 964, position 1021, or a combination thereof, as numbered accordng to SEQ ID NO: 2.

[1120] Embodiment 88: The polyribonucleotide of any one of embodiments 68-86, wherein the Plasmodium Ripr antigenic portion comprises a glutamine at position 646, position 964, position 1021, or a combination thereof, as numbered according to SEQ ID NO: 2.

[1121] Embodiment 89: The polyribonucleotide of any one of embodiments 68-86 and 88, wherein the Plasmodium Ripr antigenic portion comprises a glutamine at position 646, position 964, and position 1021, as numbered according to SEQ ID NO: 2.

[1122] Embodiment 90: The polyribonucleotide of any one of embodiments 68, 88, and 89, wherein the Plasmodium Ripr antigenic portion comprises or consists of an amino acid sequence according to SEQ ID NO: 173.

[1123] Embodiment 91: The polyribonucleotide of any one of embodiments 68-86, wherein the Plasmodium Ripr antigenic portion comprises an asparagine at position 103, position 144, position 228, position 303, position 334, position 480, position 498, position 506, position 526, position 646, position 964, position 1021, or a combination thereof, as numbered according to SEQ ID NO: 2.

[1124] Embodiment 92: The polyribonucleotide of any one of embodiments 68-86 and 91, wherein the Plasmodium Ripr antigenic portion comprises a glutamine at 103, position 144, position 228, position 303, position 334, position 480, position 498, position 506, position 526, position 646, position 964, position 1021, or a combination thereof, as numbered according to SEQ ID NO: 2.

[1125] Embodiment 93: The polyribonucleotide of any one of embodiments 68-86, 91 and 92, wherein the Plasmodium Ripr antigenic portion comprises a glutamine at position 103, position 144, position 228, position 303, position 334, position 480, position 498, position 506, position 526, position 646, position 964, and position 1021, as numbered according to SEQ ID NO: 2.

[1126] Embodiment 94: The polyribonucleotide of any one of embodiments 68-86 and 91-93, wherein the Plasmodium Ripr antigenic portion comprises or consists of an amino acid sequence according to SEQ ID NO: 172.

[1127] Embodiment 95: The polyribonucleotide of any one of embodiments 68-94, wherein the Plasmodium Ripr antigenic portion comprises a PMX cleavage site.

[1128] Embodiment 96: The polyribonucleotide of embodiment 95, wherein the PMX cleavage site comprises or consists of an amino acid sequence of SMLE.

[1129] Embodiment 97: The polyribonucleotide of embodiment 1 or 2, wherein the one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof comprise a Plasmodium TRAMP polypeptide or antigenic portion thereof.

[1130] Embodiment 98: The polyribonucleotide of embodiment 97, wherein the one or more Plasmodium Rh5 Invasion complex polypeptides or antigenic portions thereof comprise an antigenic portion of Plasmodlim TRAMP.

[1131] Embodiment 99: The polyribonucleotide of embodiment 98, wherein the Plasmodium TRAMP antigenic portion comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 179.

[1132] Embodiment 1OO: "The polyribonucleotide of embodiment 98 or 99, wherein the Plasmodlim TRAMP antigenic portion comprises or consists of an amino acid sequence according to SEQ ID NO: 179.

[1133] Embodiment 101: "The polyribonucleotide of any one of embodiments 98-100, wherein the Plasmodium TRAMP antigenic portion comprises one, two, three, four, five, six, seven, or eight N-linked glycosylation sites.

[1134] Embodiment 102: "The polyribonucleotide of embodiment 101, the Plasmodium TRAMP antigenic portion comprises an amino add substitution at one or more N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation.

[1135] Embodiment 103: "The polyribonucleotide of any one of embodiments 98-102, wherein the Plasmodium TRAMP antigenic portion comprises a substitution of NX[T/S] to QX[T/S].

[1136] Embodiment 104: The polyribonucleotide of any one of embodiments 101-103, wherein the Plasmodium TRAMP antigenic portion comprises an amino add substitution at all of the N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation.

[1137] Embodiment 105: The polyribonucleotide of any one of embodiments 98-104, wherein the Plasmodium TRAMP antigenic portion comprises an asparagine at position 149, position 195, position 202, or a combination thereof, as numbered according to SEQ ID NO: 4.

[1138] Embodiment 106: "The polyribonucleotide of any one of embodiments 98, and 101-104, wherein the Plasmodium TRAMP antigenic portion comprises a glutamine at position 149, position 195, position 202, or a combination thereof, as numbered according to SEQ ID NO: 4. [1139] Embodiment 107: "The polyribonudeotide of any one of embodiments 98, 101-104 and 106, wherein the Plasmodium TRAMP antigenic portion comprises a glutamine at position 149, position 195, and position 202, as numbered according to SEQ ID NO: 4.

[1140] Embodiment 108: "The polyribonucleotide of any one of embodiments 98, 101-104, 106 and 107, wherein the Plasmodium TRAMP antigenic portion comprises or consists of an amino add sequence according to SEQ ID NO: 181.

[1141] Embodiment 109: "The polyribonudeotide of any one of embodiments 98-103, 105, and 106, wherein the Plasmodium TRAMP antigenic portion comprises an asparagine at position 112, position 149, position 155, position 170, position 195, position 202, position 253, position 305, or a combination thereof, as numbered according to SEQ ID NO: 4.

[1142] Embodiment IIP: The polyribonucleotide of any one of embodiments 98, 101-107, and 109, wherein the Plasmodium TRAMP antigenic portion comprises a glutamine at position 112, position 149, position 155, position 170, position 195, position 202, position 253, position 305, or a combination thereof, as numbered according to SEQ ID NO: 4.

[1143] Embodiment 111: The polyribonucleotide of any one of embodiments 98, 101-107, 109, and 110, wherein the Plasmodium TRAMP antigenic portion comprises a glutamine at position 112, position 149, position 155, position 170, position 195, position 202, position 253, and position 305, as numbered according to SEQ ID NO: 4. [1144] Embodiment 112: The polyribonudeotide of any one of embodiments 98, 101-107, and 109-111, wherein the Plasmodium TRAMP antigenic portion comprises or consists of an amino add sequence according to SEQ ID NO: 180.

[1145] Embodiment 113: "The polyribonudeotide of any one of embodiments 98-112, wherein the Plasmodium TRAMP antigenic portion comprises a PMX deavage site.

[1146] Embodiment 114: The polyribonucleotide of embodiment 113, wherein the PMX deavage site comprises or consists of an amino acid sequence of HFLQ.

[1147] Embodiment 115: The polyribonucleotide of any one of embodiments 97-114, wherein the Plasmodium TRAMP polypeptide or antigenic portion thereof comprises a SUB2 deavage site.

[1148] Embodiment 116: "The polyribonucleotide of embodiment 115, wherein the SUB2 cleavage site comprises or consists of an amino add sequence according to SEQ ID NO: 193.

[1149] Embodiment 117: The polyribonucleotide of embodiment 1 or 2, wherein the one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof comprise a Plasmodium CSS polypeptide or antigenic portion thereof.

[1150] Embodiment 118: The polyribonucleotide of embodiment 117, wherein the one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof comprise an antigenic portion of Plasmodium CSS.

[1151] Embodiment 119: The polyribonudeotide of embodiment 118, wherein the antigenic portion of Plasmodium CSS comprises one or more cysteine to serine mutations.

[1152] Embodiment 120: The polyribonudeotide of embodiment 118 or 119, wherein the antigenic portion of Plasmodium CSS comprises a C30S mutation, a C80S mutation, a C276S mutation, or a combination thereof.

[1153] Embodiment 121: "The polyribonudeotide of any one of embodiments 118-120, wherein the antigenic portion of Plasmodium CSS comprises a serine at position 30 and position 80, as numbered according to SEQ ID NO: 5 or 214. [1154] Embodiment 122: The polyribonucleotide of any one of embodiments 118-121, wherein the Plasmodium CSS antigenic portion comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 194.

[1155] Embodiment 123: The polyribonucleotide of any one of embodiments 118-122, wherein the Plasmodium CSS antigenic portion comprises or consists of an amino add sequence accordng to SEQ ID NO: 194.

[1156] Embodiment 124: The polyribonudeotide of any one of embodiments 118-120, wherein the antigenic portion of Plasmodium CSS comprises a serine at position 30 and position 276, as numbered according to SEQ ID NO: 5 or 214.

[1157] Embodiment 125: The polyribonucleotide of any one of embodiments 118-120 and 124, wherein the Ptasmodum CSS antigenic portion comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 195.

[1158] Embodiment 126: The polyribonudeotide of any one of embodiments 118-120, 124, and 125, wherein the Ptasmodum CSS antigenic portion comprises or consists of an amino add sequence according to SEQ ID NO: 195.

[1159] Embodiment 127: The polyribonucleotide of any one of embodiments 118-126, wherein the Plasmodium CSS antigenic portion comprises or consists of an amino add sequence with at least 85% sequence Identity to an amino acid sequence according to SEQ ID NO: 182.

[1160] Embodiment 128: The polyribonucleotide of any one of embodiments 118-127, wherein the Plasmodium CSS antigenic portion comprises or consists of an amino add sequence according to SEQ ID NO: 182.

[1161] Embodiment 129: The polyribonudeotide of any one of embodiments 118-128, wherein the Plasmodium CSS antigenic portion comprises one, two, three, four, five, or six N-linked glycosylation sites.

[1162] Embodiment 130: The polyribonucleotide of embodiment 129, wherein the Plasmodium CSS antigenic portion comprises an amino add substitution at one or more N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation.

[1163] Embodiment 131: The polyribonucleotide of any one of embodiments 118-130, wherein the Plasmodium CSS antigenic portion comprises a substitution of NX[T/S] to QX[T/S].

[1164] Embodiment 132: The polyribonudeotide of any one of embodiments 129-131, wherein the Ptasmodum CSS antigenic portion comprises an amino add substitution at all of the N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation.

[1165] Embodiment 133: The polyribonudeotide of any one of embodiments 118-131, wherein the Ptasmodum CSS antigenic portion comprises an asparagine at position 74, position 88, or a combination thereof, as numbered according to SEQ ID NO: 5 or 214.

[1166] Embodiment 134: The polyribonudeotide of any one of embodiments 129-131 and 132, wherein the Ptasmodum CSS antigenic portion comprises a glutamine at position 74, position 88, or a combination thereof, as numbered according to SEQ ID NO: 5 or 214.

[1167] Embodiment 135: The polyribonudeotide of any one of embodiments 129-131, 132 and 134, wherein the Ptasmodum CSS antigenic portion comprises a glutamine at position 74 and position 88, as numbered according to SEQ ID NO: 5 or 214. [1168] Embodiment 136: The polyribonudeotide of any one of embodiments 129-131, 132, 134, and 135, wherein the Plasmodium CSS antigenic portion comprises or consists of an amino add sequence with at least 85% identity to an amino add sequence according to SEQ ID NO: 184.

[1169] Embodiment 137: The polyribonudeotide of any one of embodiments 129-131, 132, and 134-136, wherein the Plasmodium CSS antigenic portion comprises or consists of an amino add sequence according to SEQ ID NO: 184.

[1170] Embodiment 138: "The polyribonucleotide of any one of embodiments 118-131, wherein the Plasmodium CSS antigenic portion comprises an asparagine at position 192, position 234, position 261, position 283, or a combination thereof, as numbered according to SEQ ID NO: 195.

[1171] Embodiment 139: The polyribonucleotide of any one of embodiments 129-131, wherein the Plasmodium CSS antigenic portion comprises a glutamine at position 192, position 234, position 261, position 283, or a combination thereof, as numbered according to SEQ ID NO: 195.

[1172] Embodiment 140: The polyribonucleotide of any one of embodiments 129-131 and 139, wherein the Plasmodium CSS antigenic portion comprises a glutamine at position 192, position 234, position 261, and position 283, as numbered according to SEQ ID NO: 195.

[1173] Embodiment 141: The polyribonucleotide of any one of embodiments 118-133 or 139-140, wherein the Plasmodium CSS antigenic portion comprises or consists of an amino acid sequence according to SEQ ID NO: 185.

[1174] Embodiment 142: The polyribonucleotide of any one of embodiments 118-141, wherein the Plasmodium CSS antigenic portion comprises an asparagine at position 74, position 88, position 192, position 234, position 261, position 283, or a combination thereof, as numbered according to SEQ ID NO: 182.

[1175] Embodiment 143: The polyribonucleotide of any one of embodiments 118-142, wherein the Plasmodium CSS antigenic portion comprises a glutamine at position 74, position 88, position 192, position 234, position 261, position 283, or a combination thereof, as numbered according to SEQ ID NO: 182.

[1176] Embodiment 144: The polyribonucleotide of any one of embodiments 118-143, wherein the Plasmodium CSS antigenic portion comprises a glutamine at position 74, position 88, position 192, position 234, position 261, and position 283, as numbered according to SEQ ID NO: 182.

[1177] Embodiment 145: The polyribonucleotide of any one of embodiments 118-144, wherein the Plasmodium CSS antigenic portion comprises or consists of an amino add sequence according to SEQ ID NO: 183.

[1178] Embodiment 146: The polyribonucleotide of any one of embodiments 1-145, wherein the polypeptide comprises two or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof.

[1179] Embodiment 147: The polyribonucleotide of embodiment 146, wherein the Plasmodium Rh5 Invasion complex polypeptides or antigenic portions thereof comprise two selected from: (I) one or more Plasmodium Rh5 polypeptides or antigenic portions thereof, (ii) one or more Plasmodium CyRPA polypeptides or antigenic portions thereof, (iii) one or more Plasmodium Rfc>r polypeptides or antigenic portions thereof, (iv) one or more Plasmodium P113 polypeptides or antigenic portions thereof, (v) one or more PlasmodiumTRNAP polypeptides or antigenic portions thereof, and (vi) one or more Plasmodium CSS polypeptides or antigenic portions thereof.

[1180] Embodiment 148: The polyribonudeotide of embodiment 146 or 147, wherein the polypeptide comprises one or more Plasmodium Rh5 antigenic portions and further comprises (I) one or more Plasmodium CyRPA polypeptides or antigenic portions thereof, (ii) one or more Plasmodium Ripr polypeptides or antigenic portions thereof, (iii) one or more Plasmodium P113 polypeptides or antigenic portions thereof, (iv) one or more Plasmodium TRAMP polypeptides or antigenic portions thereof, or (v) one or more Plasmodium CSS polypeptides or antigenic portions thereof.

[1181] Embodiment 149: The polyribonucleotide of embodiment 146, wherein the polypeptide comprises:

(i) one or more Plasmodium Rh5 polypeptides or antigenic portions thereof and one or more Plasmodium CyRPA polypeptides or antigenic portions thereof;

(ii) one or more Plasmodium Rh5 polypeptides or antigenic portions thereof and one or more Plasmodium P113 polypeptides or antigenic portions thereof;

(iii) one or more Plasmodium Rh5 polypeptides or antigenic portions thereof and one or more a Plasmodium Ripr polypeptides or antigenic portions thereof;

(iv) one or more Plasmodium Rh5 polypeptides or antigenic portions thereof and one or more a Plasmodium TRAMP polypeptides or antigenic portions thereof; or

(v) one or more Plasmodium Rh5 polypeptides or antigenic portions thereof and one or more a Plasmodium CSS polypeptides or antigenic portions thereof.

[1182] Embodiment 150: The polyribonucleotide of any one of embodiments 1-149, wherein the polypeptide comprises a secretory signal.

[1183] Embodiment 151: The polyribonucleotide of embodiment 150, wherein the secretory signal comprises or consists of a heterologous secretory signal.

[1184] Embodiment 152: The polyribonucleotide of embodiment 151, wherein the heterologous secretory signal comprises or consists of a viral secretory signal.

[1185] Embodiment 153: The polyribonucleotide of embodiment 152, wherein the viral secretory signal comprises or consists of an HSV secretory signal.

[1186] Embodiment 154: The polyribonucleotide of embodiment 153, wherein the HSV secretory signal comprises or consists of an HSV-1 or HSV-2 secretory signal.

[1187] Embodiment 155: The polyribonucleotide of embodiment 153 or 154, wherein the HSV secretory signal comprises or consists of an HSV glycoprotein D (gD) secretory signal.

[1188] Embodiment 156: The polyribonucleotide of embodiment 155, wherein the HSV gD secretory signal consists of an amino add sequence accorcfing to SEQ ID NO: 42.

[1189] Embodiment 157: The polyribonudeotide of embodiment 151, wherein the secretory signal comprises or consists of a Plasmodium secretory signal.

[1190] Embodiment 158: "The polyribonucleotide of embodiment 157, wherein the secretory signal comprises or consists of a Plasmodium CyRPA secretory signal.

[1191] Embodiment 159: The polyribonudeotide of embodiment 157, wherein the secretory signal comprises or consists of a Plasmodium Ripr secretory signal.

[1192] Embodiment 160: The polyribonudeotide of embodiment 157, wherein the secretory signal comprises or consists of a Plasmodium CSS secretory signal.

[1193] Embodiment 161: The polyribonudeotide of any one of embodiments 157-160, wherein the Plasmodium secretory signal comprises or consists of an amino add sequence according to SEQ ID NOs: 186, 187, 188. [1194] Embodiment 162: The polyribonucleotide of any one of embodiments 150-161, wherein the secretory signal is located at the N-terminus of the polypeptide.

[1195] Embodiment 163: The polyribonucleotide of any one of embodiments 1-162, wherein the polypeptide comprises a transmembrane region.

[1196] Embodiment 164: The polyribonucleotide of embodiment 163, wherein the transmembrane region comprises or consists of a Plasmodium transmembrane region.

[1197] Embodiment 165: The polyribonucleotide of embodiment 164, wherein the transmembrane region comprises or consists of a P113 Plasmodium transmembrane region.

[1198] Embodiment 166: The polyribonucleotide of embodiment 164, wherein the transmembrane region comprises or consists of a TRAMP Plasmodium transmembrane region.

[1199] Embodiment 167: The polyribonucleotide of embodiment 163, wherein the transmembrane region is a heterologous transmembrane region.

[1200] Embodiment 168: The polyribonucleotide of embodiment 167, wherein the heterologous transmembrane region comprises or consists of a viral transmembrane region.

[1201] Embodiment 169: The polyribonucleotide of embodiment 168, wherein the viral transmembrane region comprises or consists of an HSV transmembrane region.

[1202] Embodiment 170: The polyribonucleotide of embodiment 169, wherein the HSV transmembrane region comprises or consists of an HSV-1 or HSV-2 transmembrane reckon.

[1203] Embodiment 171: The polyribonucleotide of embodiment 169 or 170, wherein the HSV transmembrane region comprises or consists of an HSV gD transmembrane region.

[1204] Embodiment 172: The polyribonucleotide of embodiment 171, wherein the HSV gD transmembrane region comprises or consists of an amino acid sequence according to SEQ ID NO: 75.

[1205] Embodiment 173: The polyribonucleotide of any one of embodiments 1-162, wherein the polypeptide does not comprise a transmembrane region.

[1206] Embodiment 174: The polyribonucleotide of any one of embodiments 1-173, wherein the polypeptide comprises a multimerization domain.

[1207] Embodiment 175: The polyribonucleotide of embodiment 174, wherein the multimerization domain Is a trimerization domain.

[1208] Embodiment 176: The polyribonucleotide of embodiment 175, wherein the trimerization domain is a C -terminal domain of T4 fibritin (fbldon domain).

[1209] Embodiment 177: The polyribonucleotide of embodiment 176, wherein the fbldon domain comprises or consists of an amino add sequence according to SEQ ID NO: 78.

[1210] Embodiment 178: The polyribonucleotide of any one of embodiments 1-177, wherein the polypeptide comprises a self-assembling nanopartide domain.

[1211] Embodiment 179: The polyribonucleotide of embodiment 178, wherein the self-assembling nanopartide domain is a ferritin domain.

[1212] Embodiment 180: The polyribonucleotide of embodiment 179, wherein the ferritin domain is from H. pylori.

[1213] Embodiment 181: The polyribonucleotide of embodiment 179, wherein the ferritin domain comprises or consists of a sequence according to SEQ ID NO: 88. [1214] Embodiment 182: The polyribonucleotide of any one of embodiments 1-181, wherein the polypeptide comprises one or more linkers.

[1215] Embodiment 183: "The polyribonucleotide of embodiment 182, wherein the one or more linkers comprise one or more gtydne-serine linkers.

[1216] Embodiment 184: "The polyribonucleotide of embodiment 182, wherein the one or more linkers comprise or consist of an amino acid sequence according to SEQ ID NO: 80.

[1217] Embodiment 185: The polyribonucleotide of embodiment 182, wherein the one or more linkers comprise or consist of an amino acid sequence according to SEQ ID NO: 83.

[1218] Embodiment 186: The polyribonucleotide of embodiment 182, wherein the one or more linkers comprise or consist of an amino acid sequence according to SEQ ID NO: 84.

[1219] Embodiment 187: The polyribonucleotide of embodiment 182, wherein the one or more linkers comprise or consist of an amino acid sequence according to SEQ ID NO: 86.

[1220] Embodiment 188: The polyribonucleotide of embodiment 182, wherein the one or more linkers comprise or consist of an amino add sequence according to SEQ ID NO: 137.

[1221] Embodiment 189: The polyribonucleotide of embodiment 182, wherein the one or more linkers comprise or consist of an amino add sequence according to SEQ ID NO: 138.

[1222] Embodiment 190: The polyribonucleotide of any one of embodiments 1-189, wherein the polypeptide comprises a linker after an amino add sequence of the one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof.

[1223] Embodiment 191: The polyribonucleotide of any one of embodiments 1-190, wherein Plasmodium Is

Plasmodium faldparum.

[1224] Embodiment 192: The polyribonucleotide of any one of embodiments 1-191, wherein the one or more Plasmodium Rh5 Invasion complex polypeptides or antigenic portions thereof are one or more P. faldparum Rh5 invasion complex polypeptides or antigenic portions thereof.

[1225] Embodiment 193: The polyribonucleotide of any one of embodiments 1-192, wherein the one or more Plasmodium Rh5 polypeptides or antigenic portions thereof are one or more P. fakiparum Rh5 antigenic portions.

[1226] Embodiment 194: The polyribonucleotide of any one of embodiments 1-192, wherein the one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof are a P. fakiparum CyRPA polypeptide or antigenic portion thereof.

[1227] Embodiment 195: The polyribonucleotide of any one of embodiments 1-192, wherein the one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof are a P. fakiparum P113 polypeptide or antigenic portion thereof.

[1228] Embodiment 196: The polyribonucleotide of any one of embodiments 1-192, wherein the one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof are a P. fakiparum Ripr polypeptide or antigenic portion thereof.

[1229] Embodiment 197: "The polyribonucleotide of any one of embodiments 1-192, wherein the one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof are a P. fakiparummKW polypeptide or antigenic portion thereof. [1230] Embodiment 198: "The polyribonudeotide of any one of embodiments 1-192, wherein the one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof are a P. falciparum CSS polypeptide or antigenic portion thereof.

[1231] Embodiment 199: "The polyribonucleotide of any one of embodiments 1-198, wherein Plasmodium falciparum is Plasmodium falciparum isolate 3D7.

[1232] Embodiment 200: "The polyribonucleotide of embodiment 1 or 2, wherein the polypeptide comprises (i) a secretory signal, and

(II) two Plasmodium Rh5 ordered domains.

[1233] Embodiment 201: The polyribonucleotide of embodiment 1 or 2, wherein the polypeptide comprises

(I) a secretory signal,

(II) two Plasmodium Rh5 ordered domains,

(Hi) a linker, and

(iv) a multimerization domain.

[1234] Embodiment 202: The polyribonucleotide of embodiment 1 or 2, wherein the polypeptide comprises

(I) a secretory signal,

(II) two Plasmodium Rh5 ordered domains,

(Hi) a linker, and

(iv) a transmembrane region.

[1235] Embodiment 203: The polyribonucleotide of embodiment 1 or 2, wherein the polypeptide comprises

(I) a secretory signal,

(II) two Plasmodium Rh5 ordered domains,

(HI) a linker, and

(Iv) a self-assembling nanopartide domain.

[1236] Embodiment 204: The polyribonudeotide of any one of embodiments 200-203, wherein the secretory signal is a viral secretory signal.

[1237] Embodiment 205: The polyribonucleotide of embodiment 204, wherein the secretory signal is an HSV glycoprotein D (gD) secretory signal.

[1238] Embodiment 206: The polyribonucleotide of embodiment 205, wherein the HSV gD secretory signal consists of an amino add sequence according to SEQ ID NO: 42.

[1239] Embodiment 207: The polyribonudeotide of any one of embodiments 200-206, wherein the polypeptide does not comprise a Plasmodium Rh5 disordered domain.

[1240] Embodiment 208: The polyribonudeotide of any one of embodiments 200-207, wherein the two Plasmodium Rh5 ordered domains are directly adjacent.

[1241] Embodiment 209: The polyribonudeotide of any one of embodiments 200-208, wherein the polypeptide comprises a C203Y substitution, as numbered according to SEQ ID NO: 1.

[1242] Embodiment 210: The polyribonudeotide of any one of embodiments 200-209, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 94.

[1243] Embodiment 211: The polyribonudeotide of any one of embodiments 200-210, wherein the polypeptide comprises or consists of an amino add sequence accordhg to SEQ ID NO: 94. [1244] Embodiment 212: "The polyribonudeotide of any one of embodiments 200-211, wherein the linker comprises or consists of a glycine -serine linker.

[1245] Embodiment 213: "The polyribonucleotide of any one of embodiments 200-211, wherein the linker comprises or consists of an amino acid sequence according to SEQ ID NO: 86.

[1246] Embodiment 214: "The polyribonucleotide of any one of embodiments 200-213, wherein the multimerization domain is a C-terminal domain of T4 fibritin (fbldon domain).

[1247] Embodiment 215: "The polyribonucleotide of embodiment 214, wherein the foldon domain comprises or consists of an amino add sequence according to SEQ ID NO: 78.

[1248] Embodiment 216: The polyribonucleotide of any one of embodiments 214-215, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence Identity to an amino acid sequence according to SEQ ID NO: 90.

[1249] Embodiment 217: The polyribonudeotide of any one of embodiments 214-216, wherein the polypeptide comprises or consists of an amino acid sequence according to SEQ ID NO: 90.

[1250] Embodiment 218: The polyribonudeotide of any one of embodiments 202 or 204-217, wherein the transmembrane region is a viral transmembrane region.

[1251] Embodiment 219: The polyribonucleotide of embodiment 218, wherein the viral transmembrane region Is a HSV gD transmembrane region.

[1252] Embodiment 220: The polyribonucleotide of embodiment 219, wherein the HSV gD transmembrane region comprises or consists of an amino acid sequence according to SEQ ID NO: 75.

[1253] Embodiment 221: The polyribonudeotide of any one of embodiments 218-220, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 92.

[1254] Embodiment 222: The polyribonucleotide of any one of embodiments 218-221, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 92.

[1255] Embodiment 223: The polyribonucleotide of any one of embodiments 201-203, wherein the linker comprises or consists of a glycine-serine linker.

[1256] Embodiment 224: The polyribonudeotide of any one of embodiments 201-203, wherein the linker comprises or consists of an amino add sequence according to SEQ ID NO: 137.

[1257] Embodiment 225: The polyribonudeotide of any one of embodiments 203-224, wherein the selfassembling nanoparticle domain is a ferritin domain.

[1258] Embodiment 226: The polyribonucleotide of embodiment 225, wherein the ferritin domain comprises or consists of a sequence according to SEQ ID NO: 88.

[1259] Embodiment 227: The polyribonudeotide of any one of embodiments 224-226, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 96.

[1260] Embodiment 228: The polyribonudeotide of any one of embodiments 224-227, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 96.

[1261] Embodiment 229: The polyribonudeotide of any one of embodiments 200-228, wherein the Plasmodium Rh5 ordered domains comprise one or more N-linked glycosylation sites. [1262] Embodiment 230: "The polyribonudeotide of any one of embodiments 200-229, wherein the Plasmodium Rh5 ordered domains comprise one NX[T/S] to QX[T/S] substitution.

[1263] Embodiment 231: "The polyribonucleotide of embodiment 230, wherein the Plasmodium Rh5 ordered domains comprise an N214Q substitution, as numbered according to SEQ ID NO: 1.

[1264] Embodiment 232: "The polyribonudeotide of any one of embodiments 230-231, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 99.

[1265] Embodiment 233: "The polyribonudeotide of any one of embodiments 230-231, wherein the polypeptide comprises or consists of an amino acid sequence according to SEQ ID NO: 99.

[1266] Embodiment 234: The polyribonudeotide of any one of embodiments 230-231, wherein the polypeptide comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 98.

[1267] Embodiment 235: The polyribonucleotide of any one of embodiments 230-231, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 98.

[1268] Embodiment 236: The polyribonucleotide of any one of embodiments 230-231, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 101.

[1269] Embodiment 237: The polyribonudeotide of any one of embodiments 230-231, wherein the polypeptide comprises or consists of an amino add sequence accordhg to SEQ ID NO: 101.

[1270] Embodiment 238: The polyribonudeotide of any one of embodiments 230-231, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 100.

[1271] Embodiment 239: The polyribonucleotide of any one of embodiments 230-231, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 100.

[1272] Embodiment 240: The polyribonucleotide of any one of embodiments 200-203, wherein the Plasmodium Rh5 ordered domains comprise two NX[T/S] to QX[T/S] substitutions.

[1273] Embodiment 241: The polyribonucleotide of embodiment 240, wherein the two substitutions comprise N214Q and N297Q substitutions.

[1274] Embodiment 242: The polyribonudeotide of any one of embodiments 240-241, wherein the Plasmodium Rh5 ordered domains comprise an amino add substitution at all of the N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation.

[1275] Embodiment 243: The polyribonudeotide of any one of embodiments 240-241, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 95.

[1276] Embodiment 244: The polyribonucleotide of any one of embodiments 240-241, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 95.

[1277] Embodiment 245: The polyribonudeotide of any one of embodiments 240-241, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 91. [1278] Embodiment 246: "The polyribonudeotide of any one of embodiments 240-241, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 91.

[1279] Embodiment 247: The polyribonucleotide of any one of embodiments 240-241, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 93.

[1280] Embodiment 248: "The polyribonudeotide of any one of embodiments 240-241, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 93.

[1281] Embodiment 249: "The polyribonudeotide of any one of embodiments 240-241, wherein the polypeptide comprises or consists of an amino acid sequence with at least 85% sequence Identity to an amino acid sequence according to SEQ ID NO: 97.

[1282] Embodiment 250: The polyribonucleotide of any one of embodiments 240-241, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 97.

[1283] Embodiment 251: The polyribonucleotide of embodiment 1 or 2, wherein the polypeptide comprises

(I) a secretory signal,

(II) a Plasmodium Rh5 N-terminal disordered domain,

(ill) a Plasmodium Rh5 ordered domain,

(iv) a Plasmodium Rh5 linking disordered domain, and

(v) a Plasmodium Rh5 C-terminal ordered domain.

[1284] Embodiment 252: The polyribonucleotide of embodiment 251, wherein the secretory slgial is a viral secretory signal.

[1285] Embodiment 253: The polyribonucleotide of embodiment 252, wherein the secretory signal is an HSV glycoprotein D (gD) secretory signal.

[1286] Embodiment 254: The polyribonucleotide of embodiment 253, wherein the HSV gD secretory signal consists of an amino add sequence according to SEQ ID NO: 42.

[1287] Embodiment 255: The polyribonucleotide of any one of embodiments 251-254, wherein the polypeptide comprises one or more N-llnked glycosylation sites.

[1288] Embodiment 256: The polyribonudeotide of any one of embodiments 251-255, wherein the polypeptide comprises an amino add substitution at one or more of the N-linked glycosylation sites, wherein the amino acid substitution prevents glycosylation.

[1289] Embodiment 257: The polyribonudeotide of embodiment 256, wherein the polypeptide comprises an amino acid substitution at all of the N-linked glycosylation sites, wherein the amino acid substitution prevents glycosylation.

[1290] Embodiment 258: The polyribonudeotide of any one of embodiments 251-257, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 89.

[1291] Embodiment 259: The polyribonudeotide of any one of embodiments 251-258, wherein the polypeptide is or comprises an amino add sequence according to SEQ ID NO: 89.

[1292] Embodiment 260: The polyribonudeotide of embodiment 1 or 2, wherein the polypeptide comprises (i) a secretory signal, and

(II) a Plasmodium CyRPA polypeptide or antigenic portion thereof. [1293] Embodiment 261: The polyribonucleotide of embodiment 1 or 2, wherein the polypeptide comprises

(I) a secretory signal,

(II) a Plasmodium CyRPA polypeptide or antigenic portion thereof,

(Hi) a linker, and

(iv) a multimerization domain.

[1294] Embodiment 262: The polyribonucleotide of embodiment 1 or 2, wherein the polypeptide comprises

(I) a secretory signal,

(II) a Plasmodium CyRPA polypeptide or antigenic portion thereof, and

(iii) a transmembrane region.

[1295] Embodiment 263: The polyribonucleotide of embodiment 1 or 2, wherein the polypeptide comprises

(I) a secretory signal,

(II) a Plasmodium CyRPA polypeptide or antigenic portion thereof,

(Hi) a linker, and

(iv) a transmembrane region.

[1296] Embodiment 264: The polyribonucleotide of any one of embodiments 260-263, wherein the secretory signal is a viral secretory signal.

[1297] Embodiment 265: The polyribonucleotide of embodiment 264, wherein the viral secretory signal is an HSV glycoprotein D (gD) secretory signal.

[1298] Embodiment 266: The polyribonucleotide of embodiment 265, wherein the HSV gD secretory signal comprises or consists of an amino add sequence according to SEQ ID NO: 42.

[1299] Embodiment 267: The polyribonucleotide of any one of embodiments 260-263, wherein the secretory signal is a Plasmodium secretory signal.

[1300] Embodiment 268: The polyribonucleotide of embodiment 267, wherein the Plasmodium secretory signal Is a Plasmodium secretory signal.

[1301] Embodiment 269: The polyribonucleotide of embodiment 268, wherein the Plasmodium CyRPA secretory signal comprises or consists of an amino add sequence according to SEQ ID NO: 187.

[1302] Embodiment 270: The polyribonudeotlde of any one of embodiments 260-269, wherein the Ptasmodum CyRPA polypeptide or antigenic portion thereof comprises or consists of an amino acid sequence with at least 85% sequence identity to a sequence comprising SEQ ID NO: 133.

[1303] Embodiment 271: The polyribonucleotide of any one of embodiments 260-269, wherein the Plasmodium CyRPA polypeptide or antigenic portion thereof comprises or consists of SEQ ID NO: 169 or 133.

[1304] Embodiment 272: The polyribonudeotlde of any one of embodiments 260-269, wherein the Plasmodium CyRPA polypeptide or antigenic portion thereof comprises or consists of amino acids 29-362 of SEQ ID NO: 169 or 133.

[1305] Embodiment 273: The polyribonucleotide of any one of embodiments 260-269, wherein the Ptasmodum CyRPA polypeptide or antigenic portion thereof comprises one or more N-linked glycosylation sites.

[1306] Embodiment 274: The polyribonucleotide of embodiment 273, wherein the Plasmodium CyRPA polypeptide or antigenic portion thereof comprises an amino acid substitution at one or more of the N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation. [1307] Embodiment 275: The polyribonudeotide of any one of embodiments 273-274, wherein the Plasmodium CyRPA polypeptide or antigenic portion thereof comprises a substitution of NX[T/S] to QX[T/S] and/or a substitution of NX[T/S] to NXA.

[1308] Embodiment 276: The polyribonudeotide of any one of embodiments 273-275, wherein the Plasmodium polypeptide or antigenic portion thereof comprises an N145Q, N322Q, N338Q or a combination thereof.

[1309] Embodiment 277: "The polyribonucleotide of any one of embodiments 273-276, wherein the Plasmodium CyRPK polypeptide or antigenic portion thereof comprises an N145Q, N322Q, N338Q.

[1310] Embodiment 278: The polyribonucleotide of any one of embodiments 260-277, wherein the linker comprises a glycine-serine linker.

[1311] Embodiment 279: The polyribonucleotide of embodiment 278, wherein the linker comprises or consists of an amino add sequence accordng to SEQ ID NO: 86.

[1312] Embodiment 280: The polyribonucleotide of embodiment 278, wherein the linker comprises or consists of an amino add sequence according to SEQ ID NO: 138.

[1313] Embodiment 281: The polyribonucleotide of any one of embodiments 261 or 264-280, wherein the multimerization domain is a C-terminal domain of T4 fibritin (fbldon domain).

[1314] Embodiment 282: The polyribonucleotide of embodiment 281, wherein the fbldon domain comprises or consists of an amino acid sequence according to SEQ ID NO: 78.

[1315] Embodiment 283: The polyribonucleotide of any one of embodiments 262-280, wherein the transmembrane region Is a viral transmembrane region.

[1316] Embodiment 284: The polyribonucleotide of embodiment 283, wherein the transmembrane region is a HSV gD transmembrane region.

[1317] Embodiment 285: The polyribonucleotide of embodiment 284, wherein the HSV gD transmembrane region comprises or consists of an amino add sequence according to SEQ ID NO: 75.

[1318] Embodiment 286: The polyribonucleotide of any one of embodiments 260-263, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 140.

[1319] Embodiment 287: The polyribonucleotide of any one of embodiments 260-263, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 140.

[1320] Embodiment 288: The polyribonucleotide of any one of embodiments 260-263, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 141.

[1321] Embodiment 289: The polyribonucleotide of any one of embodiments 260-263, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 141.

[1322] Embodiment 290: The polyribonucleotide of any one of embodiments 260-263, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 142.

[1323] Embodiment 291: The polyribonucleotide of any one of embodiments 260-263, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 142. [1324] Embodiment 292: "The polyribonudeotide of any one of embodiments 260-263, wherein the polypeptide comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 143.

[1325] Embodiment 293: "The polyribonucleotide of any one of embodiments 260-263, wherein the polypeptide comprises or consists of an amino acid sequence according to SEQ ID NO: 143.

[1326] Embodiment 294: "The polyribonucleotide of any one of embodiments 260-263, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 144.

[1327] Embodiment 295: The polyribonucleotide of any one of embodiments 260-263, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 144.

[1328] Embodiment 296: The polyribonucleotide of any one of embodiments 260-263, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 198.

[1329] Embodiment 297: The polyribonudeotide of any one of embodiments 260-263, wherein the polypeptide comprises or consists of an amino acid sequence according to SEQ ID NO: 198.

[1330] Embodiment 298: The polyribonudeotide of any one of embodiments 260-263, wherein the polypeptide comprises or consists of an amino add sequence with at lead: 85% sequence Identity to an amino add sequence according to SEQ ID NO: 199.

[1331] Embodiment 299: The polyribonudeotide of any one of embodiments 260-263, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 199.

[1332] Embodiment 300: The polyribonucleotide of embodiment 1 or 2, wherein the polypeptide comprises

(I) a secretory signal,

(II) a Plasmodium Pl 13 polypeptide or antigenic portion thereof, and

(III) a transmembrane region.

[1333] Embodiment 301: The polyribonucleotide of embodiment 1 or 2, wherein the polypeptide comprises

(I) a secretory signal,

(II) a Plasmodium Pl 13 polypeptide or antigenic portion thereof,

(iii) a linker, and

(iv) a transmembrane region.

[1334] Embodiment 302: The polyribonucleotide of embodiment 300 or 301, wherein the secretory signal is a viral secretory signal.

[1335] Embodiment 303: The polyribonucleotide of embodiment 302, wherein the secretory signal is an HSV glycoprotein D (gD) secretory signal.

[1336] Embodiment 304: The polyribonudeotide of embodiment 303, wherein the HSV gD secretory signal consists of an amino add sequence according to SEQ ID NO: 42.

[1337] Embodiment 305: The polyribonudeotide of any one of embodiments 301-304, wherein the linker comprises a glycine-serine linker.

[1338] Embodiment 306: The polyribonudeotide of any one of embodiments 301-304, wherein the linker comprises or consists of an amino add sequence according to SEQ ID NO: 138. [1339] Embodiment 307: "The polyribonucleotide of any one of embodiments 300-306, wherein the transmembrane region is a viral transmembrane region.

[1340] Embodiment 308: "The polyribonucleotide of embodiment 307, wherein the viral transmembrane region is a HSV gD transmembrane region.

[1341] Embodiment 309: "The polyribonucleotide of embodiment 308, wherein the HSV gD transmembrane region comprises or consists of an amino add sequence according to SEQ ID NO: 75.

[1342] Embodiment 310: "The polyribonucleotide of any one of embodiments 300-309, wherein the Plasmodium P113 polypeptide or antigenic portion thereof comprises or consists of an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 135.

[1343] Embodiment 311: "The polyribonucleotide of any one of embodiments 300-309, wherein the Plasmodium P113 polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 135.

[1344] Embodiment 312: The polyribonucleotide of any one of embodiments 300-311, wherein the Plasmodium Pl 13 polypeptide or antigenic portion thereof comprises one or more N-llnked glycosylation sites.

[1345] Embodiment 313: The polyribonucleotide of embodiment 312, wherein the Plasmodium P113 polypeptide or antigenic portion thereof comprises an amino acid substitution at one or more of the N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation.

[1346] Embodiment 314: The polyribonucleotide of any one of embodiments 300-313, wherein the Plasmodium Pl 13 polypeptide or antigenic portion thereof comprises one or more substitutions of NX[T/S] to QX[T/S] and/or substitutions of NX[T/S] to NXA.

[1347] Embodiment 315: The polyribonucleotide of any one of embodiments 300-313, wherein the Plasmodium Pl 13 antigenic portion comprises an asparagine at 207, position 268, position 317, position 360, position 661, position 697, position 779, position 876, or a combination thereof, as numbered according to SEQ ID NO: 6.

[1348] Embodiment 316: The polyribonudeotide of any one of embodiments 300-314, wherein the Plasmodium P113 antigenic portion comprises a glutamine at position 207, position 268, position 317, position 360, position 661, position 697, position 779, position 876, or a combination thereof, as numbered according to SEQ ID NO: 6.

[1349] Embodiment 317: "The polyribonucleotide of any one of embodiments 300-314, wherein the Plasmodium P113 antigenic portion comprises a glutamine position 207, position 268, position 317, position 360, position 661, position 697, position 779, and position 876, as numbered according to SEQ ID NO: 6.

[1350] Embodiment 318: "The polyribonucleotide of any one of embodiments 300-314 or 316-317, wherein the Plasmodium Pl 13 polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 136.

[1351] Embodiment 319: "The polyribonucleotide of any one of embodiments 300-314 or 316-317, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 196.

[1352] Embodiment 320: "The polyribonucleotide of any one of embodiments 300-314 or 316-317, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 196. [1353] Embodiment 321: The polyribonucleotide of any one of embodiments 300-314 or 316-317, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 197.

[1354] Embodiment 322: "The polyribonucleotide of any one of embodiments 300-314 or 316-317, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 197.

[1355] Embodiment 323: "The polyribonucleotide of embodiment 1 or 2, wherein the polypeptide comprises (i) a secretory signal, and

(II) a Plasmodium Ripr polypeptide or antigenic portion thereof.

[1356] Embodiment 324: The polyribonucleotide of embodiment 1 or 2, wherein the polypeptide comprises

(i) a secretory signal,

(II) a Plasmodium Ripr polypeptide or antigenic portion thereof, and

(iii) a transmembrane region.

[1357] Embodiment 325: The polyribonucleotide of embodiment 1 or 2, wherein the polypeptide comprises

(I) a secretory signal,

(II) a Plasmodium Ripr polypeptide or antigenic portion thereof,

(Hi) a linker, and

(iv) a transmembrane region.

[1358] Embodiment 326: The polyribonucleotide of any one of embodiments 323-325, wherein the secretory signal is a Plasmodium secretory signal.

[1359] Embodiment 327: The polyribonucleotide of embodiment 326, wherein the Plasmodium secretary signal is a Plasmodium Ripr secretory signal.

[1360] Embodiment 328: The polyribonucleotide of embodiment 327, wherein the Plasmodium Ripr secretory signal comprises or consists of an amino acid sequence according to SEQ ID NO: 186.

[1361] Embodiment 329: The polyribonucleotide of any one of embodiments 323-325, wherein the secretory signal is a viral secretory signal.

[1362] Embodiment 330: The polyribonucleotide of embodiment 329, wherein the secretory signal is an HSV glycoprotein D (gD) secretory signal.

[1363] Embodiment 331: The polyribonucleotide of embodiment 330, wherein the HSV gD secretory signal comprises or consists of an amino add sequence according to SEQ ID NO: 42.

[1364] Embodiment 332: The polyribonucleotide of any one of embodiments 323-331, wherein the Plasmodium Ripr polypeptide or antigenic portion thereof Ripr comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 171.

[1365] Embodiment 333: The polyribonucleotide of any one of embodiments 323-331, wherein the Plasmodium Ripr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 171.

[1366] Embodiment 334: The polyribonucleotide of any one of embodiments 323-325, wherein the Plasmodium Ripr polypeptide or antigenic portion thereof comprises one, two, three, four, five, six, seven, eight, nine, ten, eleven or twelve N-linked glycosylation sites. [1367] Embodiment 335: "The polyribonucleotide of embodiment 334, wherein the Plasmodium Ripr polypeptide or antigenic portion thereof comprises an amino acid substitution at one or more N-linked glycosylation sites, wherein the amino acid substitution prevents glycosylation.

[1368] Embodiment 336: "The polyribonucleotide of any one of embodiments 334-335, wherein the Plasmodium Ripr polypeptide or antigenic portion thereof comprises a substitution of NX[T/S] to QX[T/S].

[1369] Embodiment 337: "The polyribonucleotide of any one of embodiments 334-336, wherein the Plasmodium Ripr polypeptide or antigenic portion thereof comprises an amino add substitution at all of the N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation.

[1370] Embodiment 338: The polyribonucleotide of any one of embodiments 334-335, wherein the Plasmodium Ripr polypeptide or antigenic portion thereof comprises an asparagine at position 103, position 144, position 228, position 303, position 334, position 480, position 498, position 506, position 526, position 646, position 964, position 1021, or a combination thereof, as numbered according to SEQ ID NO: 171.

[1371] Embodiment 339: The polyribonucleotide of any one of embodiments 334-337, wherein the Plasmodium Ripr polypeptide or antigenic portion thereof comprises a glutamhe at position 103, position 144, position 228, position 303, position 334, position 480, position 498, position 506, position 526, position 646, position 964, position 1021, or a combination thereof, as numbered according to SEQ ID NO: 171.

[1372] Embodiment 340: The polyribonucleotide of any one of embodiments 334-337, wherein the Plasmodium Ripr polypeptide or antigenic portion thereof comprises a glutamhe at position 103, position 144, position 228, position 303, position 334, position 480, position 498, position 506, position 526, position 646, position 964, and position 1021, as numbered according to SEQ ID NO: 171.

[1373] Embodiment 341: The polyribonucleotide of any one of embodiments 334-337 or 339-340, wherein the Plasmodium Ripr polypeptide or antigenic portion thereof comprises or consists of an amino acid sequence according to SEQ ID NO: 172.

[1374] Embodiment 342: The polyribonucleotide of any one of embodiments 325-341, wherein the linker comprises a glycine-serine linker.

[1375] Embodiment 343: The polyribonucleotide of any one of embodiments 325-342, wherein the linker comprises or consists of an amino acid sequence according to SEQ ID NO: 138.

[1376] Embodiment 344: The polyribonucleotide of any one of embodiments 323-325, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 145.

[1377] Embodiment 345: The polyribonudeotide of any one of embodiments 323-325, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 145.

[1378] Embodiment 346: The polyribonudeotide of any one of embodiments 323-325, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 147.

[1379] Embodiment 347: The polyribonudeotide of any one of embodiments 323-325, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 147.

[1380] Embodiment 348: The polyribonudeotide of any one of embodiments 323-325, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 149. [1381] Embodiment 349: "The polyribonudeotide of any one of embodiments 323-325, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 149.

[1382] Embodiment 350: "The polyribonucleotide of any one of embodiments 323-325, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 151.

[1383] Embodiment 351: "The polyribonucleotide of any one of embodiments 323-325, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 151.

[1384] Embodiment 352: "The polyribonucleotide of any one of embodiments 324-351, wherein the transmembrane region is a heterologous transmembrane region.

[1385] Embodiment 353: The polyribonucleotide of embodiment 352, wherein the heterologous transmembrane region comprises or consists of a viral transmembrane region.

[1385] Embodiment 354: The polyribonucleotide of embodiment 352, wherein the heterologous transmembrane region comprises or consists of an HSV transmembrane region.

[1387] Embodiment 355: The polyribonucleotide of embodiment 354, wherein the HSV transmembrane region comprises or consists of an HSV gD transmembrane region.

[1388] Embodiment 356: The polyribonucleotide of embodiment 355, wherein the HSV gD transmembrane region comprises or consists of an amino acid sequence according to SEQ ID NO: 75.

[1389] Embodiment 357: The polyribonucleotide of any one of embodiments 323-325, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 146.

[1390] Embodiment 358: The polyribonucleotide of any one of embodiments 323-325, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 146.

[1391] Embodiment 359: The polyribonucleotide of any one of embodiments 323-325, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 200.

[1392] Embodiment 360: The polyribonucleotide of any one of embodiments 323-325, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 200.

[1393] Embodiment 361: The polyribonucleotide of any one of embodiments 323-325, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 148.

[1394] Embodiment 362: The polyribonudeotide of any one of embodiments 323-325, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 148.

[1395] Embodiment 363: The polyribonudeotide of any one of embodiments 323-325, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 201.

[1396] Embodiment 364: The polyribonudeotide of any one of embodiments 323-325, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 201.

[1397] Embodiment 365: The polyribonudeotide of any one of embodiments 323-325, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 150. [1398] Embodiment 366: The polyribonucleotide of any one of embodiments 323-325, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 150.

[1399] Embodiment 367: "The polyribonucleotide of any one of embodiments 323-325, wherein the polypeptide comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 202.

[1400] Embodiment 368: "The polyribonucleotide of any one of embodiments 323-325, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 202.

[1401] Embodiment 369: The polyribonucleotide of any one of embodiments 323-325, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 152.

[1402] Embodiment 370: The polyribonucleotide of any one of embodiments 323-325, wherein the polypeptide comprises or consists of an amino acid sequence according to SEQ ID NO: 152.

[1403] Embodiment 371: The polyribonucleotide of any one of embodiments 323-325, wherein the polypeptide comprises or consists of an amino acid sequence with at least 85% sequence Identity to an amino acid sequence according to SEQ ID NO: 203.

[1404] Embodiment 372: The polyribonucleotide of any one of embodiments 323-325, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 203.

[1405] Embodiment 373: The polyribonucleotide of any one of embodiments 323-325, wherein the Plasmodium Ripr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 178.

[1406] Embodiment 374: The polyribonucleotide of any one of embodiments 323-325, wherein the Plasmodium Ripr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 177.

[1407] Embodiment 375: The polyribonucleotide of any one of embodiments 323-325, wherein the Plasmodium Ripr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 177.

[1408] Embodiment 376: The polyribonucleotide of any one of embodiments 323-325, wherein the Plasmodium Ripr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 176.

[1409] Embodiment 377: The polyribonucleotide of any one of embodiments 323-325, wherein the Plasmodium Ripr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 176.

[1410] Embodiment 378: The polyribonucleotide of any one of embodiments 323-325, wherein the Plasmodium Ripr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 175.

[1411] Embodiment 379: The polyribonucleotide of any one of embodiments 323-325, wherein the Plasmodium Ripr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 175. [1412] Embodiment 380: "The potyribonucleotide of any one of embodiments 323-325, wherein the Plasmodium Rjpr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 174.

[1413] Embodiment 381: "The polyribonucleotide of any one of embodiments 323-325, wherein the Plasmodium Rjpr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 174.

[1414] Embodiment 382: "The polyribonucleotide of any one of embodiments 323-325, wherein the Plasmodium Rjpr polypeptide or antigenic portion thereof comprises or consists of an amino acid sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 190.

[1415] Embodiment 383: The polyribonucleotide of any one of embodiments 323-325, wherein the Plasmodium Ripr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 190.

[1416] Embodiment 384: The polyribonucleotide of any one of embodiments 323-383, wherein the Plasmodium Ripr polypeptide or antigenic portion thereof comprises an amino acid substitution at one or more N- linked glycosylation sites, wherein the amino add substitution prevents glycosylation.

[1417] Embodiment 385: The potyribonucleotide of any one of embodiments 323-384, wherein the Plasmodium Ripr polypeptide or antigenic portion thereof comprises a substitution of NX[T/S] to QX[T/S].

[1418] Embodiment 386: The polyribonucleotide of any one of embodiments 323-385, wherein the Plasmodium Ripr polypeptide or antigenic portion thereof comprises an amino acid substitution at all of the N-llnked glycosylation sites, wherein the amino add substitution prevents glycosylation.

[1419] Embodiment 387: The potyribonucleotide of any one of embodiments 323-383, wherein the Plasmodium Ripr polypeptide or antigenic portion thereof comprises an asparagine at position 646, position 964, position 1021, or a combination thereof, as numbered according to SEQ ID NO: 190.

[1420] Embodiment 388: The polyribonudeotide of any one of embodiments 323-386, wherein the Plasmodium Ripr polypeptide or antigenic portion thereof comprises a glutamine at position 646, position 964, position 1021, or a combination thereof, as numbered according to SEQ ID NO: 190.

[1421] Embodiment 389: The polyribonudeotide of any one of embodiments 323-386, wherein the Piasmodum Ripr polypeptide or antigenic portion thereof comprises a glutamine at position 646, position 964, and position 1021, as numbered according to SEQ ID NO: 190.

[1422] Embodiment 390: The polyribonucleotide of any one of embodiments 323-389, wherein the Piasmodum Ripr polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 173.

[1423] Embodiment 391: The polyribonudeotide of any one of embodiments 323-390, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 153.

[1424] Embodiment 392: The polyribonudeotide of any one of embodiments 323-390, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 153.

[1425] Embodiment 393: The polyribonudeotide of any one of embodiments 323-390, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 204. [1426] Embodiment 394: "The potyribonucleotide of any one of embodiments 323-390, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 204.

[1427] Embodiment 395: "The polyribonucleotide of any one of embodiments 323-390, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 154.

[1428] Embodiment 396: "The polyribonucleotide of any one of embodiments 323-390, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 154.

[1429] Embodiment 397: "The polyribonucleotide of any one of embodiments 323-390, wherein the polypeptide comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 205.

[1430] Embodiment 398: The polyribonucleotide of any one of embodiments 323-390, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 205.

[1431] Embodiment 399: The polyribonucleotide of any one of embodiments 323-390, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence Identity to an amino acid sequence according to SEQ ID NO: 155.

[1432] Embodiment 400: The potyribonucleotide of any one of embodiments 323-390, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 155.

[1433] Embodiment 401: The polyribonucleotide of any one of embodiments 323-390, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 206.

[1434] Embodiment 402: The polyribonucleotide of any one of embodiments 323-390, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 206.

[1435] Embodiment 403: The polyribonucleotide of any one of embodiments 323-390, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 156.

[1436] Embodiment 404: The polyribonucleotide of any one of embodiments 323-390, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 156.

[1437] Embodiment 405: The polyribonucleotide of any one of embodiments 323-390, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 207.

[1438] Embodiment 406: The polyribonucleotide of any one of embodiments 323-390, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 207.

[1439] Embodiment 407: The polyribonucleotide of any one of embodiments 323-390, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 157.

[1440] Embodiment 408: The polyribonucleotide of any one of embodiments 323-390, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 157.

[1441] Embodiment 409: The potyribonucleotide of any one of embodiments 323-390, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 208. [1442] Embodiment 410: "The potyribonucleotide of any one of embodiments 323-390, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 208.

[1443] Embodiment 411: "The polyribonucleotide of any one of embodiments 323-390, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 158.

[1444] Embodiment 412: "The polyribonucleotide of any one of embodiments 323-390, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 158.

[1445] Embodiment 413: "The polyribonucleotide of any one of embodiments 323-390, wherein the polypeptide comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 209.

[1446] Embodiment 414: The polyribonucleotide of any one of embodiments 323-390, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 209.

[1447] Embodiment 415: The polyribonucleotide of embodiment 1 or 2, wherein the polypeptide comprises

(I) a secretory signal,

(II) a Plasmodium TRAMP polypeptide or antigenic portion thereof.

[1448] Embodiment 416: The polyribonucleotide of embodiment 415, wherein the secretory signal is a viral secretory signal.

[1449] Embodiment 417: The polyribonucleotide of embodiment 416, wherein the viral secretory signal Is an HSV glycoprotein D (gD) secretory signal.

[1450] Embodiment 418: The polyribonucleotide of embodiment 417, wherein the HSV gD secretory signal comprises or consists of an amino add sequence according to SEQ ID NO: 42.

[1451] Embodiment 419: The potyribonucleotide of any one of embodiments 415-418, wherein the Plasmodium TRAMP polypeptide or antigenic portion thereof comprises or consists of an amino acid sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 179.

[1452] Embodiment 420: The polyribonucleotide of any one of embodiments 415-419, wherein the Plasmodium TRAMP polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 179.

[1453] Embodiment 421: The polyribonucleotide of any one of embodiments 415-418, wherein the Plasmodium TRAMP polypeptide or antigenic portion thereof comprises one, two, three, four, five, six, seven, or eight N-inked glycosylation sites.

[1454] Embodiment 422: The polyribonucleotide of any one of embodiments 415-421, wherein the Plasmodium TRAMP polypeptide or antigenic portion thereof comprises an amino add substitution at one or more N- linked glycosylation sites, wherein the amino add substitution prevents glycosylation.

[1455] Embodiment 423: The polyribonucleotide of any one of embodiments 415-422, wherein the Plasmodium TRAMP polypeptide or antigenic portion thereof comprises a substitution of NX[T/S] to QX[T/S],

[1456] Embodiment 424: The polyribonucleotide of embodiment 421, wherein the Plasmodium TRAMP polypeptide or antigenic portion thereof comprises an amino add substitution at all of the N-finked glycosylation sites, wherein the amino acid substitution prevents glycosylation. [1457] Embodiment 425: The polyribonudeotide of any one of embodiments 415-423, wherein the Plasmodium TRAMP polypeptide or antigenic portion thereof comprises an asparagine at position 149, position 195, position 202, or a combination thereof, as numbered according to SEQ ID NO: 179.

[1458] Embodiment 426: The polyribonudeotide of any one of embodiments 415-424, wherein the Plasmodium TRAMP polypeptide or antigenic portion thereof comprises a glutamine at position 149, position 195, position 202, or a combination thereof, as numbered according to SEQ ID NO: 179.

[1459] Embodiment 427: The polyribonudeotide of embodiment 426, wherein the PlasmodiumTRAMP polypeptide or antigenic portion thereof comprises a glutamine at position 149, position 195, and position 202, as numbered according to SEQ ID NO: 179.

[1460] Embodiment 428: The polyribonudeotide of any one of embodiments 415-418 and 421-427, wherein the Plasmodium TRAMP polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 181.

[1461] Embodiment 429: The polyribonucleotide of any one of embodiments 415-423, wherein the Plasmodium TRAMP polypeptide or antigenic portion thereof comprises an asparagine at position 112, position 149, position 155, position 170, position 195, position 202, position 253, position 305, or a combination thereof, as numbered according to SEQ ID NO: 179.

[1462] Embodiment 430: The polyribonucleotide of any one of embodiments 415-424 and 426-429, wherein the Plasmodium TRAMP polypeptide or antigenic portion thereof comprises a glutamine at position 112, position 149, position 155, position 170, position 195, position 202, position 253, position 305, or a combination thereof, as numbered according to SEQ ID NO: 179.

[1463] Embodiment 431: The polyribonucleotide of any one of embodiments 415-424 and 426-430, wherein the Plasmodium TRAMP polypeptide or antigenic portion thereof comprises a glutamine at position 112, position 149, position 155, position 170, position 195, position 202, position 253, and position 305, as numbered according to SEQ ID NO: 179.

[1464] Embodlment 432: The polyribonucleotide of any one of embodiments 415-424 and 426-431, wherein the Plasmodium TRAMP polypeptide or antigenic portion thereof comprises or consists of an amino add sequence according to SEQ ID NO: 180.

[1465] Embodiment 433: The polyribonucleotide of any one of embodiments 415-432, wherein the Plasmodium TRAMP polypeptide or antigenic portion thereof comprises a PMX deavage site.

[1466] Embodiment 434: The polyribonucleotide of embodiment 433, wherein the PMX deavage site comprises or consists of an amino add sequence of HFLQ.

[1467] Embodiment 435: The polyribonucleotide of any one of embodiments 415-434, wherein the Plasmodium TRAMP polypeptide or antigenic portion thereof comprises a SLIB2 deavage site.

[1468] Embodiment 436: The polyribonucleotide of embodiment 435, wherein the SUB2 cleavage site comprises or consists of an amino add sequence according to SEQ ID NO: 193.

[1469] Embodiment 437: The polyribonucleotide of any one of embodiments 415-436, wherein the linker comprises a glycine-serine linker.

[1470] Embodiment 438: The polyribonudeotide of any one of embodiments 415-437, wherein the linker comprises or consists of an amino add sequence according to SEQ ID NO: 138. [1471] Embodiment 439: The polyribonucleotide of embodiment 415, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 159.

[1472] Embodiment 440: The polyribonucleotide of embodiment 415, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 159.

[1473] Embodiment 441: The polyribonucleotide of embodiment 415, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 160.

[1474] Embodiment 442: The polyribonucleotide of embodiment 415, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 160.

[1475] Embodiment 443: The polyribonucleotide of embodiment 415, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 161.

[1476] Embodiment 444: The polyribonucleotide of embodiment 415, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 161.

[1477] Embodiment 445: The polyribonucleotide of embodiment 1 or 2, wherein the polypeptide comprises

(I) a secretory signal, and

(II) a Plasmodium CSS polypeptide or antigenic portion thereof.

[1478] Embodiment 446: The polyribonucleotide of embodiment 1 or 2, wherein the polypeptide comprises

(I) a secretory signal,

(II) a Plasmodium CSS polypeptide or antigenic portion thereof, and

(III) a transmembrane region.

[1479] Embodiment 447: The polyribonucleotide of embodiment 1 or 2, wherein the polypeptide comprises

(I) a secretory signal,

(II) a Plasmodium CSS polypeptide or antigenic portion thereof,

(III) a linker, and

(iv) a transmembrane region.

[1480] Embodiment 448: The polyribonucleotide of any one of embodiments 445-447, wherein the secretory signal is a Plasmodium secretory signal.

[1481] Embodiment 449: The polyribonucleotide of embodiment 448, wherein the Plasmodium secretory signal is a Plasmodium CSS secretory signal.

[1482] Embodiment 450: The polyribonucleotide of embodiment 449, wherein the Plasmodium CSS secretory signal comprises or consists of an amino add sequence according to SEQ ID NO: 188.

[1483] Embodiment 451: The polyribonucleotide of any one of embodiments 445-447, wherein the secretory signal is a viral secretory signal.

[1484] Embodiment 452: The polyribonucleotide of embodiment 451, wherein the viral secretory signal is an

HSV glycoprotein D (gD) secretory signal.

[1485] Embodiment 453: The polyribonucleotide of embodiment 452, wherein the HSV gD secretory signal comprises or consists of an amino add sequence according to SEQ ID NO: 42. [1486] Embodiment 454: The polyribonucleotide of any one of embodiments 445-453, wherein the Plasmodium CSS polypeptide or antigenic portion thereof comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 182.

[1487] Embodiment 455: The polyribonucleotide of any one of embodiments 445-454, wherein the Plasmodium CSS polypeptide or antigenic portion thereof comprises or consists of an amino acid sequence according to SEQ ID NO: 182.

[1488] Embodiment 456: The polyribonucleotide of any one of embodiments 445-453, wherein the Plasmodium CSS polypeptide or antigenic portion thereof comprises one, two, three, four, five, or six N-linked glycosylation sites.

[1489] Embodiment 457: The polyribonucleotide of any one of embodiments 445-456, wherein the Plasmodium CSS polypeptide or antigenic portion thereof comprises an amino acid substitution at one or more N- linked glycosylation sites, wherein the amino add substitution prevents glycosylation.

[1490] Embodiment 458: The polyribonucleotide of any one of embodiments 445-457, wherein the Plasmodium CSS polypeptide or antigenic portion thereof comprises a substitution of NX[T/S] to QX[T/S].

[1491] Embodiment 459: The polyribonucleotide of any one of embodiments 456-458, wherein the Plasmodium CSS polypeptide or antigenic portion thereof comprises an amino add substitution at all of the N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation.

[1492] Embodiment 460: The polyribonucleotide of any one of embodiments 445-459, wherein the Plasmodium CSS polypeptide or antigenic portion thereof comprises an asparagine at position 74, position 88, position 192, position 234, position 261, and position 283, or a combination thereof, as numbered according to SEQ ID NO: 182.

[1493] Embodiment 461: "The polyribonucleotide of any one of embodiments 445-459, wherein the Plasmodium CSS polypeptide or antigenic portion thereof comprises a glutamine at position 74, position 88, position 192, position 234, position 261, position 283, or a combination thereof, as numbered according to SEQ ID NO: 182.

[1494] Embodiment 462: The polyribonucleotide of any one of embodiments 445-459 and 461, wherein the Plasmodium CSS polypeptide or antigenic portion thereof comprises a glutamine at 74, position 88, position 192, position 234, position 261, and position 283, as numbered according to SEQ ID NO: 182.

[1495] Embodiment 463: The polyribonucleotide of any one of embodiments 445-459, 461 and 462, wherein the Plasmodium CSS polypeptide or antigenic portion thereof comprises or consists of an amino acid sequence according to SEQ ID NO: 183.

[1496] Embodiment 464: The polyribonucleotide of any one of embodiments 445-459, wherein the linker comprises a glycine-serine linker.

[1497] Embodiment 465: The polyribonucleotide of any one of embodiments 445-459, wherein the linker comprises or consists of an amino acid sequence according to SEQ ID NO: 138.

[1498] Embodiment 466: The polyribonucleotide of any one of embodiments 445-459, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 162.

[1499] Embodiment 467: The polyribonucleotide of any one of embodiments 445-447, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 162. [1500] Embodiment 468: "The polyribonucleotide of any one of embodiments 445-447, wherein the polypeptide comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 163.

[1501] Embodiment 469: "The polyribonucleotide of any one of embodiments 445-447, wherein the polypeptide comprises or consists of an amino acid sequence according to SEQ ID NO: 163.

[1502] Embodiment 470: "The polyribonucleotide of any one of embodiments 445-447, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 164.

[1503] Embodiment 471: The polyribonucleotide of any one of embodiments 445-447, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 164.

[1504] Embodiment 472: The polyribonucleotide of any one of embodiments 445-453, wherein the Plasmodium CSS polypeptide or antigenic portion thereof comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 194.

[1505] Embodiment 473: The polyribonucleotide of any one of embodiments 445-453 and 472, wherein the Plasmodium CSS polypeptide or antigenic portion thereof comprises or consists of an amino acid sequence according to SEQ ID NO: 194.

[1506] Embodiment 474: The polyribonucleotide of any one of embodiments 445-453, wherein the Plasmodium CSS polypeptide or antigenic portion thereof comprises or consists of an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 195.

[1507] Embodiment 475: The polyribonudeotlde of any one of embodiments 445-453 and 474, wherein the Plasmodium CSS polypeptide or antigenic portion thereof comprises or consists of an amino acid sequence according to SEQ ID NO: 195.

[1508] Embodiment 476: The polyribonucleotide of any one of embodiments 445-453, wherein the Plasmodium CSS polypeptide or antigenic portion thereof comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 182.

[1509] Embodiment 477: The polyribonudeotlde of any one of embodiments 445-453 and 476, wherein the Plasmodium CSS polypeptide or antigenic portion thereof comprises or consists of an amino acid sequence according to SEQ ID NO: 182.

[1510] Embodiment 478: The polyribonudeotlde of any one of embodiments 472-477, wherein the Plasmodium CSS polypeptide or antigenic portion thereof comprises one, two, three, four, five, or six N-linked glycosylation sites.

[1511] Embodiment 479: The polyribonudeotlde of any one of embodiments 472-478, wherein the Plasmodium CSS polypeptide or antigenic portion thereof comprises an amino acid substitution at one or more N- linked glycosylation sites, wherein the amino add substitution prevents glycosylation.

[1512] Embodiment 480: The polyribonudeotlde of any one of embodiments 472-479, wherein the Plasmodium CSS polypeptide or antigenic portion thereof comprises a substitution of NX[T/S] to QX[T/S].

[1513] Embodiment 481: The polyribonudeotlde of any one of embodiments 478-480, wherein the Plasmodium CSS polypeptide or antigenic portion thereof comprises an amino acid substitution at all of the N-linked glycosylation sites, wherein the amino add substitution prevents glycosylation. [1514] Embodiment 482: The polyribonucleotide of any one of embodiments 472-480, wherein the Plasmodium CSS polypeptide or antigenic portion thereof comprises an asparagine at position 74, position 88, or a combination thereof, as numbered according to SEQ ID NO: 194.

[1515] Embodiment 483: The polyribonucleotide of any one of embodiments 472-480, wherein the Plasmodium CSS polypeptide or antigenic portion thereof comprises a glutamine at position 74, position 88, or a combination thereof, as numbered according to SEQ ID NO: 194.

[1516] Embodiment 484: The polyribonucleotide of any one of embodiments 472-480 and 483, wherein the Plasmodium CSS polypeptide or antigenic portion thereof comprises a glutamine at position 74, and position 88, as numbered according to SEQ ID NO: 194.

[1517] Embodiment 485: The polyribonucleotide of any one of embodiments 472-480, wherein the Plasmodium CSS polypeptide or antigenic portion thereof comprises or consists of an amino acid sequence according to SEQ ID NO: 184.

[1518] Embodiment 486: The polyribonucleotide of any one of embodiments 472-480 and 485, wherein the Plasmodium CSS polypeptide or antigenic portion thereof comprises an asparagine at position 192, position 234, position 261, position 283, or a combination thereof, as numbered according to SEQ ID NO: 195.

[1519] Embodiment 487: The polyribonucleotide of any one of embodiments 472-480 and 485, wherein the Plasmodium CSS polypeptide or antigenic portion thereof comprises a glutamine at position 192, position 234, position 261, position 283, or a combination thereof, as numbered according to SEQ ID NO: 195.

[1520] Embodiment 488: The polyribonucleotide of any one of embodiments 472-480, 485, and 487, wherein the PlasmodkmiCSS polypeptide or antigenic portion thereof comprises a glutamine at position 192, position 234, position 261, and position 283, as numbered according to SEQ ID NO: 195.

[1521] Embodiment 489: The polyribonucleotide of any one of embodiments 486-488, wherein the Plasmodium CSS polypeptide or antigenic portion thereof comprises or consists of an amino acid sequence according to SEQ ID NO: 185.

[1522] Embodiment 490: The polyribonucleotide of any one of embodiments 472-489, wherein the Plasmodium CSS polypeptide or antigenic portion thereof comprises an asparagine at position 74, position 88, position 192, position 234, position 261, position 283, or a combination thereof, as numbered according to SEQ ID NO: 182.

[1523] Embodiment 491: The polyribonucleotide of any one of embodiments 472-489, wherein the Plasmodium CSS polypeptide or antigenic portion thereof comprises a glutamine at position 74, position 88, position 192, position 234, position 261, position 283, or a combination thereof, as numbered according to SEQ ID NO: 182.

[1524] Embodiment 492: The polyribonucleotide of any one of embodiments 472-489 and 491, wherein the Plasmodium CSS polypeptide or antigenic portion thereof comprises a glutamine at position 74, position 88, position 192, position 234, position 261, and position 283, as numbered according to SEQ ID NO: 182.

[1525] Embodiment 493: The polyribonucleotide of any one of embodiments 472-489, 491 and 492, wherein the Plasmodium CSS polypeptide or antigenic portion thereof comprises or consists of an amino acid sequence according to SEQ ID NO: 183.

[1526] Embodiment 494: The polyribonucleotide of any one of embodiments 472-493, wherein the first linker and/or second linker comprises or consists of an amino add sequence according to SEQ ID NO: 138. [1527] Embodiment 495: "The polyribonucleotide of any one of embodiments 472-494, wherein the transmembrane region is a heterologous transmembrane region.

[1528] Embodiment 496: "The polyribonucleotide of embodiment 495, wherein the heterologous transmembrane region comprises or consists of a viral transmembrane region.

[1529] Embodiment 497: "The polyribonucleotide of embodiment 496, wherein the viral transmembrane region comprises or consists of an HSV transmembrane region.

[1530] Embodiment 498: "The polyribonucleotide of embodiment 497, wherein the HSV transmembrane region comprises or consists of an HSV gD transmembrane region.

[1531] Embodiment 499: The polyribonucleotide of embodiment 498, wherein the HSV gD transmembrane region comprises or consists of an amino acid sequence according to SEQ ID NO: 75.

[1532] Embodiment 500: The polyribonucleotide of any one of embodiments 484-505, wherein the linker comprises a glycine-serine linker.

[1533] Embodiment 501: The polyribonucleotide of embodiment 500, wherein the linker comprises or consists of an amino add sequence according to SEQ ID NO: 138.

[1534] Embodiment 502: The polyribonucleotide of embodiment 500 or 501, wherein the polypeptide comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 165.

[1535] Embodiment 503: The polyribonucleotide of any one of embodiments 500-502, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 165.

[1536] Embodiment 504: The polyribonucleotide of embodiment 500 or 501, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 166.

[1537] Embodiment 505: The polyribonucleotide of any one of embodiments 500-502, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 166.

[1538] Embodiment 506: The polyribonucleotide of embodiment 500 or 501, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 167.

[1539] Embodiment 507: The polyribonucleotide of any one of embodiments 500-502, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 167.

[1540] Embodiment 508: The polyribonucleotide of embodiment 500 or 501, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 168.

[1541] Embodiment 509: The polyribonucleotide of any one of embodiments 500-501, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 168.

[1542] Embodiment 510: The polyribonucleotide of embodiment 500 or 501, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 210.

[1543] Embodiment 511: The polyribonucleotide of any one of embodiments 500-502, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 210. [1544] Embodiment 512: "The polyribonucleotide of embodiment 500 or 501, wherein the polypeptide comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 211.

[1545] Embodiment 513: "The polyribonucleotide of any one of embodiments 500-502, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 211.

[1546] Embodiment 514: "The polyribonucleotide of embodiment 500 or 501, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 212.

[1547] Embodiment 515: The polyribonucleotide of any one of embodiments 500-502, wherein the polypeptide comprises or consists of an amino add sequence according to SEQ ID NO: 212.

[1548] Embodiment 516: The polyribonucleotide of embodiment 500 or 501, wherein the polypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 213.

[1549] Embodiment 517: The polyribonucleotide of any one of embodiments 500-502, wherein the polypeptide comprises or consists of an amino acid sequence according to SEQ ID NO: 213.

[1550] Embodiment 518: The polyribonucleotide of any one of embodiments 1-517, wherein the polyribonucleotide Is an Isolated polyribonucleotide.

[1551] Embodiment 519: The polyribonucleotide of any one of embodiments 1-518, wherein the polyribonucleotide Is an engineered polyribonucleotide.

[1552] Embodiment 520: The polyribonucleotide of any one of embodiments 1-519, wherein the polyribonucleotide Is a codon-optimized polyribonucleotide.

[1553] Embodiment 521: An RNA construct comprising In 5' to 3' order:

(I) a S' UTR;

(ID a polyribonucleotide of any one of embodiments 1-520;

(HD a 3' UTR; and

(iv) a polyA tail sequence.

[1554] Embodiment 522: The RNA construct of embodiment 521, wherein

(i) the 5' UTR comprises or consists of a modified human alpha-globin S'-UTR; and

(ii) the 3' UTR that comprises or consists of a first sequence from the amino terminal enhancer of split (AES) messenger RNA and a second sequence from the mitochondrial encoded 12S ribosomal RNA.

[1555] Embodiment 523: The RNA construct of embodiment 521 or 522, wherein the 5' UTR consists of a ribonucleic add sequence according to SEQ ID NO: 111.

[1556] Embodiment 524: The RNA construct of any one of embodiments 521-523, wherein the 3' UTR consists of a ribonudeic add sequence according to SEQ ID NO: 117.

[1557] Embodiment 525: The RNA construct of any one of embodiments 521-524, wherein the polyA tail sequence is a split polyA tail sequence.

[1558] Embodiment 526: The RNA construct of embodiment 525, wherein the split polyA tail sequence consists of a ribonudeic add sequence according to SEQ ID NO: 114.

[1559] Embodiment 527: The RNA construct of any one of embodiments 521-526, further comprising a 5' cap. [1560] Embodiment 528: "The RNA construct of any one of embodiments 521-527, further comprising a cap proximal sequence comprising positions +1, +2, +3, +4, and +5 of the polyribonucleotide.

[1561] Embodiment 529: "The RNA construct of embodiment 527 or 528, wherein the 5' cap comprises or consists of m7(3'OMeG)(5')ppp(5')(2^,OMeAi)pG2, wherein Ai is position +1 of the polyribonucleotide, and Gz is position +2 of the polyribonucleotide.

[1562] Embodiment 530: "The RNA construct of embodiment 528 or 529, wherein the cap proximal sequence comprises Ai and Gz of the Capl structure, and a sequence comprising: A3A4U5 (SEQ ID NO: 138) at positions +3, +4 and +5 respectively of the polyribonucleotide.

[1563] Embodiment 531: The RNA construct of any one of embodiments 521-530, wherein the polyribonucleotide Includes modified uridines in place of all uridines.

[1564] Embodiment 532: The RNA construct of embodiment 522, wherein modified uridines are each Nl- methyl-pseudouridine.

[1565] Embodiment 533: A composition comprising one or more polyribonucleotides of any one of embodiments 1-520.

[1566] Embodiment 534: A composition comprising one or more RNA constructs of any one of embodiments

521-532.

[1567] Embodiment 535: The composition of embodiment 533 or 534, wherein the composition further comprises lipid nanopartides, polyplexes (PLX), lipidated polyplexes (LPLX), or liposomes, wherein the one or more polyribonucleotides are fully or partially encapsulated within the lipid nanopartides, polyplexes (PLX), lipidated polyplexes (LPLX), or liposomes.

[1568] Embodiment 536: The composition of any one of embodiments 533-535, wherein the composition further comprises lipid nanopartides, wherein the one or more polyribonucleotides are encapsulated within the lipid nanopartides.

[1569] Embodiment 537: A pharmaceutical composition comprising the composition of any one of embodiments 545-548 and at least one pharmaceutically acceptable exdpient

[1570] Embodiment 538: The pharmaceutical composition of embodiment 537, wherein the pharmaceutical comprises a cryoprotectant

[1571] Embodiment 539: The pharmaceutical composition of embodiment 537 or 538, wherein the pharmaceutical comprises an aqueous buffered solution.

[1572] Embodiment 540: The pharmaceutical composition of embodiment 539, wherein the aqueous buffered solution comprises one or more of Tris base, Tris HQ, NaCI, KCI, NazHPOi, and KH2PO4.

[1573] Embodiment 541: A combination comprising:

(i) a first pharmaceutical composition comprising a first pdyribonudeotide, wherein the first polyribonucleotide encodes a first polypeptide, and the first polypeptide comprises one or more Plasmodium Rh5 polypeptides or antigenic portions thereof; and

(ii) a second pharmaceutical composition comprising a second polyribonucleotide, wherein the second polyribonucleotide encodes a second polypeptide, the second polypeptide comprises one or more Plasmodium Rh5 invasion complex polypeptides selected from: one or more CyRPA polypeptides or antigenic portions thereof, one or more Ripr polypeptides or antigenic portions thereof, one or more P113 polypeptides or antigenic portions thereof, one or more TRAMP polypeptides or antigenic portions thereof, one or more CSS polypeptides or antigenic portions thereof, or a combination thereof.

[1574] Embodiment 542: The combination of embodiment 541, wherein the first polyribonucleotide is a polyribonucleotide according to any one of embodiments 1-37 and 200-259.

[1575] Embodiment 543: The combination of embodiment 541 or 542, wherein the second polypeptide comprises one or more one or more CyRPA polypeptides or antigenic portions thereof.

[1576] Embodiment 544: The combination of embodiment 543, wherein the second polyribonucleotide is a polyribonucleotide according to any one of embodiments 38-53 and 260-311.

[1577] Embodiment 545: The combination of embodiment 541 or 543, wherein the second polypeptide comprises one or more one or more Pl 13 polypeptides or antigenic portions thereof.

[1578] Embodiment 546: The combination of embodiment 545, wherein the second polyribonucleotide is a polyribonucleotide according to any one of embodiments 54-66 and 300-322.

[1579] Embodiment 547: The combination of embodiment 541 or 542, wherein the second polypeptide comprises one or more one or more Ripr polypeptides or antigenic portions thereof.

[1580] Embodiment 548: The combination of embodiment 547, wherein the second polyribonucleotide is a polyribonucleotide according to any one of embodiments 67-96 and 323-414.

[1581] Embodiment 549: The combination of embodiment 541 or 542, wherein the second polypeptide comprises one or more one or more TRAMP polypeptides or antigenic portions thereof.

[1582] Embodiment 550: The combination of embodiment 549, wherein the second polyribonucleotide is a polyribonucleotide according to any one of embodiments 97-116 and 415-444.

[1583] Embodiment 551: The combination of embodiment 541 or 542, wherein the second polypeptide comprises one or more one or more CSS polypeptides or antigenic portions thereof.

[1584] Embodiment 552: The combination of embodiment 551, wherein the second polyribonucleotide is a polyribonucleotide according to any one of embodiments 117-145 and 415-517.

[1585] Embodiment 553: A combination comprising:

(i) a first pharmaceutical composition conprising a first polyribonucleotide of any one of embodiments 1- 520, wherein the first polyribonucleotide encodes a first polypeptide, and the first polypeptide comprises one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof; and

(ii) a second pharmaceutical composition comprising a second polyribonucleotide, wherein the second polyribonucleotide encodes a second polypeptide, the second polypeptide comprises one or more Plasmocfiumt cell antigens.

[1586] Embodiment 554: A combination comprising:

(I) a first pharmaceutical composition comprising a first polyribonucleotide of any one of embodiments 1- 520, wherein the first polyribonucleotide encodes a first polypeptide, and the first polypeptide comprises one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof; and (ii) a second pharmaceutical composition comprising a second polyribonucleotide, wherein the second polyribonucleotide encodes a second polypeptide, the second polypeptide comprises one or more Plasmodium CSP polypeptides or antigenic portions thereof. [1587] Embodiment 555: A method comprising administering a polyribonucleotide according to any one of embodiments 1-520 to a subject

[1588] Embodiment 556: A method comprising administering an RNA construct according to any one of embodiments 521-532 to a subject

[1589] Embodiment 557: A method comprising administering a composition according to any one of embodiments 533-536 to a subject.

[1590] Embodiment 558: A method comprising administering one or more doses of the pharmaceutical composition of any one of embodiments 537-540 to a subject.

[1591] Embodiment 559: The pharmaceutical composition of any one of embodiments 537-540 for use in the treatment of a malaria Infection comprising administering one or more doses of the pharmaceutical composition to a subject

[1592] Embodiment 560: The pharmaceutical composition of any one of embodiments 537-540 for use in the prevention of a malaria infection comprising administering one or more doses of the pharmaceutical composition to a subject.

[1593] Embodiment 561: The method of any one of embodiments 555-558 or the pharmaceutical composition for use of embodiment 571 or 572, comprising administering two or more doses of the pharmaceutical composition to a subject.

[1594] Embodiment 562: The method of any one of embodiments 555-558 and 561 or the pharmaceutical composition for use of any one of embodiments 559-561, comprising administering three or more doses of the pharmaceutical composition to a subject.

[1595] Embodiment 563: A method comprising administering a combination of any one of embodiments 541-554 to a subject

[1596] Embodiment 564: The method of embodiment 563, wherein the first pharmaceutical composition and the second pharmaceutical composition are administered on the same day.

[1597] Embodiment 565: The method of embodiment 563 and 564, wherein the first pharmaceutical composition and the second pharmaceutical composition are administered on different days.

[1598] Embodiment 566: The method of any one of embodiments 563-565, wherein the first pharmaceutical composition and the second pharmaceutical composition are administered to the subject at different locations on the subject's body.

[1599] Embodiment 567: The method of any one of embodiments 555-558 and 561-566, wherein the method is a method of treating a malaria infection.

[1600] Embodiment 568: The method of any one of embodiments 555-558 and 561-567, wherein the method is a method of preventing a malaria infection.

[1601] Embodiment 569: The method of any one of embodiments 555-558 and 561-568, wherein the subject has or is at risk of developing a malaria infection.

[1602] Embodiment 570: The method of any one of embodiments 555-558 and 561-569, wherein the subject is a human.

[1603] Embodiment 571: The method of any one of embodiments 555-558 and 561-570, wherein administration induces an anti-malaria Immune response in the subject [1604] Embodiment 572: "The method of embodiment 571, wherein the anti-malaria immune response in the subject comprises an adaptive immune response.

[1605] Embodiment 573: "The method of embodiment 572, wherein the anti -malaria immune response in the subject comprises a T-cell response.

[1606] Embodiment 574: The method of embodiment 573, wherein the T-cell response is or comprises a CD4+ T cell response.

[1607] Embodiment 575: "The method of embodiment 573, wherein the T-cell response is or comprises a

CD8+ T cell response.

[1608] Embodiment 576: The method of any one of embodiments 571-575, wherein the anti-malaria Immune system response comprises a B-cell response.

[1609] Embodiment 577: The method of any one of embodiments 571-576, wherein the anti-malaria immune system response comprises the production of antibodies directed against the one or more Plasmodium antigens.

[1610] Embodiment 578: Use of the pharmaceutical composition of any one of embodiments 537-540 in the treatment of a malaria infection.

[1611] Embodiment 579: Use of the pharmaceutical composition of any one of embodiments 537-540 in the prevention of a malaria Infection.

[1612] Embodiment 580: Use of the pharmaceutical composition of any one of embodiments 537-540 In inducing an anti-malaria immune response in a subject.

[1613] Embodiment 581: A polypeptide encoded by a polyribonucleotide of any one of embodiments 1-520.

[1614] Embodiment 582: A polypeptide encoded by an RNA construct of any one of embodiments 521-532.

[1615] Embodiment 583: A host cell comprising a polyribonucleotide of any one of embodiments 1-520.

[1616] Embodiment 584: A host cell comprising an RNA construct of any one of embodiments 521-532.

[1617] Embodiment 585: A host cell comprising a polypeptide of embodiment 581 or 582.

EXEMPLIFICATION

Example 1: In-vitro Expression of Exemplary Polyribonucleotides Encoding Plasmodium Polypeptide

Constructs

[1618] The present Example describes In vitro expression of exemplary polyribonucleotides encoding different malarial Rh5 invasion complex antigens, as described herein.

[1619] In vitro expression assays can be used to assess expression and localization (e.g., intracellular, surface) of different Plasmodium polypeptide constructs In mammalian cells (e.g., HEK293T cells). Various malarial Rh5 invasion complex polypeptide constructs are provided herein. For example, various polypeptide constructs are provided In Section IIH above, amino add sequences for exemplary Rh5 polypeptide constructs are provided In Table 7, amino acid sequences for exemplary CyRPA polypeptide constructs are provided in Table 10, amino add sequences for exemplary Ripr polypeptide constructs are provided in Table 13, amino acid sequences for exemplary TRAMP polypeptide constructs are provided in Table 16, amino add sequences for exemplary CSS polypeptide constructs are provided In Table 19, and amino add sequences for exemplary GARP polypeptide constructs are provided in Table 22. [1620] Assays can be initially performed with non-fbrmulated RNA constructs to determine functionality. Formulated RNA constructs can also be assessed.

[1621] Exemplary mammalian cells, HEK293T cells, are transfected with (i) 350 ng of non-fbrmulated RNA constructs or (II) 5 ng or 50 ng of LNP formulated RNA constructs. HEK293T cells transfected with 350 ng of non- fbrmulated RNA constructs or with 5 ng of formulated RNA constructs will be assessed for protein expression by antibody staining and FACS. The transfection rate is determined for both non-fbrmulated and formulated RNA constructs. Transfection rate can be determined by measuring percentage of positive cells, and total expression can be determined by measuring median fluorescence of the total HEK population. HEK293T cells transfected with 50 ng of formulated RNA constructs can be assessed for protein secretion by detecting protein in the culture supernatant of transfected cells.

[1622] In some embodiments, an expression product of a malarial Rh5 invasion complex polypeptide construct (e.g., a Rh5 invasion complex antigen) can be detected using a suitable commercially available antibody. In some instances where there are limited resources (e.g., no available antfoodies) for an expression product of a malarial Rh5 invasion complex polypeptide construct (e.g., a Rh5 invasion complex antigen), a tagging methodology may be employed to determine expression, such as, e.g., FLAG or HiBIt etc. Corresponding antibodies specific to for a respective tag, or in the case of HiBIt - a luminescence-based assay, can be used to determine expression.

[1623] In one experiment, 400,000 HEK293T cells were seeded per well of 12 well plate. Six hours later 125ng of each RNA construct was added per well In the presence of the working concentration of commercial transfection reagent Messenger Max. After incubation for 18 hours, cells were resuspended, stained with monoclonal antibodies to RH5 and then analyzed by flow cytometry to determine surface expression (cells stained without permeabilization) and total expression (cells stained after permeabilization) of the RH5 constructs.

[1624] As depicted in FIGS. 14A-14F cells were Incubated with a viability dye prior to splitting for either, permeabiization and staining for intracellular protein (e.g., IC, total), or staining without permeabilization to detect surface protein (e.g., SF). More specifically, A) viability of cells treated for Intracellular staining were unaffected by transfection with 125ng of RH5 RNA constructs, 8) viability of cells treated for surface staining were unaffected by transfection with 125ng of RH5 RNA constructs, C) percentage of cells staining positive for RH5 protein by the intracellular method is shown, D) percentage of cells staining positive for RH5 protein by the surface staining method is shown, E) mean fluorescence intensity of total cellular RH5 protein is shown, and F) mean fluorescence intensity of surface localized RH5 protein is shown.

[1625] As depicted in FIGS. 15A-15D, cells were incubated with a viability dye prior to splitting for either, permeabiization and staining for intracellular protein (e.g., IC, total), or staining without permeabilization to detect surface protein (e.g., SF). In particular, A) percentage of cells staining positive for RH5 protein by the Intracellular method, B) percentage of cels staining positive for RH5 protein by the surface staining method, C) mean fluorescence intensity of total cellular RH5 protein, and D) mean fluorescence intensity of surface localized RH5 protein are shown. Figures 15A-15D utilized commercially avaiable anti-RHS antibody 9AD4 and QA1.

[1626] As depicted in figures 16A-16B, cells were incubated with a viability dye. Specifically, A) percentage of cels staining positive for a protein (e.g., Rh5 invasion complex polypeptide(s), Rh5, CyRPA, Ripr, CSS, PTRAMP, or GARP) encoded by a RNA construct, and B) mean fluorescence intensity of total protein encoded by a RNA construct are shown. [1627] As depicted in FIGS. 17A-17J, RNA constructs were tagged with a HiBiT tag and protein expression assessed using a luminescence based assay. Cellular protein expression and exported protein expression were assessed. Cells were transfected with IDng/lOOuL well of a 96 well plate in triplicate. Alter inatation for 18 hours the media was transferred to another 96 weH plate and the media replaced. HiBit reagents were added to the plates containing cells to determine the amount of protein expression (either surface bound or intracellular protein). HiBit reagents were also added to the plates containing the transferred media to determine the amount of protein exported out of the cell (e.g., not membrane bound). FIGS. 17A-17B depict the quantification of cellular and exported protein for GARP constructs, FIGS. 17C-17D depict the quantification of cellular and exported protein for CyRPA constructs, FIGS. 17E-17F depict the quantification of cellular and exported protein for Rlpr constructs, FIGS. 17G-17H depict the quantification of cellular and exported protein for CSS constructs, and FIGS. 171- 17J depict the quantification of cellular and exported protein for TRAMP constructs.

Example 2: Immunooenidtv Studies of Exemplary Polyribonucleotides Encoding Plasmodium Dolvoeotide constructs

[1628] The present Example describes an assessment of the ability of certain polyribonucleotides encoding different malarial Rh5 Invasion complex antigens, provided by the present disclosure, to induce immune responses, as assessed in mice.

[1629] C57BL6 female mice (10-12-weeks old) are Immunized intramuscularly (IM) twice, for example, on days 0 and 21, with 1 pg of a formulated RNA construct (described in Example 1) or Injected with a control (e.g., 1 pg of a formulated RNA construct which lacks a start codon) (e.g., n=8 mke/group). Blood samples are collected preImmunization (day 0) and after the first dose (e.g., on days 7, 14, 21, 28 and 35) to generate serum samples at various time points. At the end of the experiment (e.g., day 35), blood was harvested and as needed cryopreserved.

[1630] Serum samples obtained from each group of Immunized animals will be analyzed by one or more of the following method(s): (1) Enzyme-linked Immunosorbent Assay (EUSA), (2) FACS analysis of infected red blood cells, (3) growth Inhibition assay, and/or (4) complement deposition assay.

(1) Enzyme-linked Immunosorbent Assay (ELISA)

[1631] Provided polyribonucleotides can be assessed for their ability to induce production of antibodies that may bind to target Rh5 Invasion complex antigens using EUSA. RNA constructs (as described in Example 1), can be assessed for their ability to induce production of antibodies that bind to a respective target antigen (e.g., one or more portions of an associated Rh5 invasion complex polypeptide).

[1632] Briefly, MaxiSorp 96-well plates will be coated with recombinantly expressed Rh5 invasion complex antigens in coating buffer (50mM sodium carbonate, pH 9.6) and incubated overnight at 4°C. Plates will then be blocked with 1% BSA in PBS either for Ih at 37°C, or overnight at 4°C. Bound IgG will be detected using horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG. Signal will be detected after adding the substrate 3, 3', 5,5'- Tetramethylbenzidine (TMB) and 25% sulfuric add to stop the reaction. Optical densities (OD) will be read at 450 nm. Slitable commercially available antibodies will be established as positive controls.

(2) FACs Assay

[1633] Where reagents (e.g., antibodies) are unavailable to establish EUSAs, provided polyribonucleotides can be assessed for their ability to induce production of antibodies that may bind to target Rh5 invasion complex antigens by FACs. Binding of antibodies present in the sera of immunized mice will be evaluated by staining infected RBCs with sera and analysing stained samples by flow cytometry. (3) IVE using cognate immune serum

[1634] Where reagents are unavailable, immune serum from immunized mice can be used in place of monoclonal antibodies in an In Vitro Expression style assay, to determine whether an immune response against a construct was elicited in vivo. Cells are transfected with a construct and immune serum from mice immunized with the same construct respectively were used to stain the transfected cells, with untransfected cells as a negative control. When a construct elicited an immune response, expression of the construct can be detected by flow cytometry.

(4) Growth Inhibition Assay

[1635] Provided polyribonucleotides can be assessed for their ability to induce production of antibodies that Inhibit parasite blood stage growth. In some embodiments, a provided polyribonucleotide is determined to induce a useful immune response if serum from a subject (e.g., a mouse) immunized with such construct is shown to inhibit Plasmodium falciparum (Pf) blood stage growth using assay as described herein.

[1636] The impact of antibodies in immune sera on parasite blood stage growth will be tested with a growth Inhibition assay (GIA). Briefly, synchronized cultures of blood stage parasites will be setup in multi-well (e.g., 96, or 384 well plates) at 0.4% parasitemia, 2% haematocrit in a 50-10ul uL volume. Purified IgG from Immunized mice at lOmg/mL is added to duplicate wells at a highest concentration of 2mg (making up 20% of the culture), with a two fold dilution to make 1, 0.5, 0.25 and 0.125mg/mL concentrations of IgG. After Incubation at 37C for ~40 hours, parasite grown for all conditions can be determined by a variety of methods; for transgenic parasites expressing luciferase, luminescence based readout can be used, for non-transgenlc parasite strains parasite growth can be determined by intercalation of SYBR green or the Lactate Dehydrogenase (LDH) assay which is a colorimetric assay. The relative abundance of LDH in parasites results in a blue colour forming after addition of assay reagents which when read at 650nm can be used as a proxy for parasitemia. Control samples will be stained with SYBR green and samples analyzed by FACS to determine growth relative to Oh control. Experimental groups will be analyzed at 48hrs to determine whether serum inhibited growth.

(5) Complement deposition assay

[1637] The ability of antibodies present in the serum of immunized animals to activate the classical complement cascade will be quantification of Clq deposition. In brief, 96 well plates coated with purified merozoltes or target merozoite antigens will be inciiiated with immune serum, after removal and washing steps the plates will be incubated with human serum containing active complement. After washing of plates, Clq will be detected with Clq specific monoclonal antibodies followed by species relevant secondary antibodies conjugated to HRP. Signal will be detected after adding the substrate 3,3',5,5'-Tetramethyfoenzidine (TMB) and 25% sulfuric add to stop the reaction. Optical densities (OD) will be read at 450 nm. Suitable commercially available antibodies will be established as positive controls.

EQUIVALENTS

[1638] Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of technologies described herein. The scope of the present dsdosure Is not Intended to be limited to the above Description, but rather Is as set forth In the following dalms

Claims

1. A polyribonudeotide encoding a polypeptide, wherein the polypeptide comprises one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof.

2. The polyribonudeotide of daim 1, wherein the one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof comprise:

(ii) one or more /Vasma/Ajm Cysteine-Rich Protective Antigen (CyRPA) polypeptides or antigenic portions thereof;

(iv) one or more Plasmodium P113 polypeptides or antigenic portions thereof;

3. The polyribonudeotide of daim 2, wherein the one or more Plasmodium Rh5 antigenic portions comprise:

(i) an amino add sequence according to SEQ ID NO: 118, 119, 124, or 126,

(II) an amino add sequence according to SEQ ID NO: 120 or 128, or

(III) a combination thereof.

4. The polyribonudeotide of daim 2, wherein the one or more Plasmodium Rh5 antigenic portions comprise:

(I) an amino add sequence according to SEQ ID NO: 121, 122, 125, or 127, (ii) an amino acid sequence according to SEQ ID NO: 123 or 129, or (Hi) a combination thereof.

5. The polyribonudeotide of any one of claims 1-2, wherein the one or more Plasmodium Rh5 antigenic portions comprise:

(i) an amino add sequence according to SEQ ID NO: 124, 125, 126, or 127,

(ii) an amino acid sequence according to SEQ ID NO: 128 or 129, or

(III) a combination thereof.

6. The polyribonudeotide of daim 2, wherein the one or more Plasmodium Rh5 antigenic portions comprise:

(i) an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 130,

(ii) an amino acid sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 118, 119, 121, 122, 124, 125, 126, or 127,

(iii) an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 131, (iv) an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 120, 123, 128, or 129, or

(v) a combination thereof.

7. The polyribonucleotide of any one of claims 1-2, wherein the polypeptide comprises an amino add sequence with at least 85% sequence identity to an amino add sequence of positions 26-363 of SEQ ID NO: 94, 95 or 99.

8. The polyribonudeotide of any one of claims 1-2, wherein the polypeptide comprises an amino add sequence with at least 85% sequence identity to an amino add sequence of positions 26-526 of SEQ ID NO: 89.

9. The polyribonudeotide of claim 1 or 2, wherein the one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof comprise a Plasmodium CyRPA polypeptide or antigenic portion thereof.

10. The polyribonudeotide of dalm 1 or 2, wherein the one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof comprise a Plasmodium P113 polypeptide or antigenic portions thereof.

11. The polyribonudeotide of dalm 1 or 2, wherein the one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof comprise a Plasmodium Ripr polypeptide or antigenic portion thereof.

12. The polyribonudeotide of dalm 1 or 2, wherein the one or more Plasmodium Rh5 Invasion complex polypeptides or antigenic portions thereof comprise a Plasmodium TRAMP polypeptide or antigenic portion thereof.

13. The polyribonudeotide of dalm 1 or 2, wherein the one or more Plasmodium Rh5 Invasion complex polypeptides or antigenic portions thereof comprise a Plasmodium CSS polypeptide or antigenic portion thereof.

14. The polyribonudeotide of dalm 1 or 2, wherein the polypeptide comprises

(I) a secretory signal, and

(II) two Piasmodum Rh5 ordered domains.

15. The polyribonudeotide of dalm 1 or 2, wherein the polypeptide comprises

(I) a secretory signal,

(II) two Plasmodium Rh5 ordered domains,

(iii) a linker, and

(iv) a multimerization domain.

16. The polyribonudeotide of dalm 1 or 2, wherein the polypeptide comprises

(I) a secretory signal,

(II) two Piasmodum Rh5 ordered domains,

(iii) a linker, and

(Iv) a transmembrane region.

17. The pdyribonudeotide of daim 1 or 2, wherein the polypeptide comprises

(i) a secretory signal,

(ii) two Plasmodium Rh5 ordered domains,

(iii) a linker, and

(iv) a self-assembling nanopartide domain.

18. The pdyribonudeotide of any one of daims 14-17, wherein the polypeptide comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 94.

19. The pdyribonudeotide of any one of daims 14-17, wherein the pdypeptide comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 90.

20. The pdyribonudeotide of any one of daims 14-17, wherein the polypeptide comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 92.

21. The pdyribonudeotide of any one of daims 14-17, wherein the pdypeptide comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 96.

22. The pdyribonudeotide of any one of daims 14-17, wherein the pdypeptide comprises or consists of an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 99.

23. The pdyribonudeotide of any one of daims 14-17, wherein the pdypeptide comprises or consists of an amino acid sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 98.

24. The pdyribonudeotide of any one of daims 14-17, wherein the pdypeptide comprises or consists of an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 101.

25. The pdyribonudeotide of any one of daims 14-17, wherein the pdypeptide comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 100.

26. The pdyribonudeotide of any one of daims 14-17, wherein the pdypeptide comprises or consists of an amino add sequence with at least 85% sequence Identity to an amino add sequence according to SEQ ID NO: 95.

27. The pdyribonudeotide of any one of daims 14-17, wherein the pdypeptide comprises or consists of an amino add sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 91.

28. The pdyribonudeotide of any one of claims 14-17, wherein the pdypeptide comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 93.

29. The polyribonucleotide of any one of claims 14-17, wherein the polypeptide comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino acid sequence according to SEQ ID NO: 97.

30. The polyribonucleotide of claim 1 or 2, wherein the polypeptide comprises

(i) a secretory signal,

(ii) a Plasmodium Rh5 N-terminal disordered domain,

(iii) a Plasmodium Rh5 ordered domain,

(iv) a Plasmodium Rh5 linking disordered domain, and

(v) a Plasmodium Rh5 C-terminal ordered domain.

31. The polyribonucleotide of claim 30, wherein the polypeptide comprises or consists of an amino acid sequence with at least 85% sequence identity to an amino add sequence according to SEQ ID NO: 89.

32. An RNA construct comprising in 5* to 3' order:

(0 a 5' UTR;

(ii) a polyribonucleotide of any one of claims 1-532;

(Hi) a 3' UTR; and

(iv) a polyA tail sequence.

33. A composition comprising one or more polyribonucleotides of any one of claims 1-31.

34. A composition comprising one or more RNA constructs of claim 32.

35. The composition of claim 33 or 34, wherein the composition further comprises lipid nanopartides, polyplexes (PLX), lipidated polyplexes (LPLX), or liposomes, wherein the one or more polyribonucleotides are fully or partially encapsulated within the lipid nanoparticles, polyplexes (PLX), lipidated polyplexes (LPLX), or liposomes.

36. A pharmaceutical composition comprising the composition of claim 35 and at least one pharmaceutically acceptable excipient.

37. A combination comprising:

(ii) a second pharmaceutical composition comprising a second polyribonucleotide, wherein the second polyribonucleotide encodes a second polypeptide, the second polypeptide comprises one or more Plasmodium Rh5 invasion complex polypeptides selected from: one or more CyRPA polypeptides or antigenic portions thereof, one or more Ripr polypeptides or antigenic portions thereof, one or more Pl 13 polypeptides or antigenic portions thereof, one or more TRAMP polypeptides or antigenic portions thereof, one or more CSS polypeptides or antigenic portions thereof, or a combination thereof.

38. A combination comprising:

(i) a first pharmaceutical composition comprising a first polyribonucleotide polyribonucleotide of any one of claims 1-31, wherein the first polyribonucleotide encodes a first polypeptide, and the first polypeptide comprises one or more Piasmtxfium Rh5 invasion complex polypeptides or antigenic portions thereof; and

(II) a second pharmaceutical composition comprising a second polyribonucleotide, wherein the second polyribonucleotide encodes a second polypeptide, the second polypeptide comprises one or more PlasmodiumT cell antigens.

39. A combination comprising:

(i) a first pharmaceutical composition comprising a first polyribonucleotide of any one of claims 1-31, wherein the first polyribonucleotide encodes a first polypeptide, and the first polypeptide comprises one or more Plasmodium Rh5 invasion complex polypeptides or antigenic portions thereof; and

(il) a second pharmaceutical composition comprising a second polyribonucleotide, wherein the second polyribonucleotide encodes a second polypeptide, the second polypeptide comprises one or more Plasmodium C.SP polypeptides or antigenic portions thereof.

40. A method comprising administering a polyribonucleotide according to any one of claims 1-31 to a subject

41. A method comprising administering an RNA construct according to claim 32 to a subject.

42. A method comprising administering a composition according to any one of claims 33-35 to a subject.

43. A method comprising administering one or more doses of the pharmaceutical composition of claim 36 to a subject

44. The pharmaceutical composition of claim 36 for use in the treatment of a malaria infection comprising administering one or more doses of the pharmaceutical composition to a subject

45. The pharmaceutical composition of claim 36 for use in the prevention of a malaria infection comprising administering one or more doses of the pharmaceutical composition to a subject

46. A method comprising administering a combination of any one of claims 37-39 to a subject.

47. Use of the pharmaceutical composition of claim 36 in the treatment of a malaria infection.

48. Use of the pharmaceutical composition of claim 36 in the prevention of a malaria infection.

49. Use of the pharmaceutical composition of claim 36 in inducing an anti-malaria immune response in a subject

50. A polypeptide encoded by a polyribonucleotide of any one of claims 1-31.

51. A polypeptide encoded by an RNA construct of claim 32.

52. A host cell comprising a polyribonucleotide of any one of claims 1-31.

53. A host cell comprising an RNA construct of claim 32.

54. A host cell comprising a polypeptide of claim 50 or 51.