JPWO2021142458A5 - - Google Patents

Claims (15)

a bacterium engineered to have at least one expression cassette encoding a eukaryotic translatable mRNA comprising a prokaryotic promoter, an internal ribosome entry sequence (IRES) , and a nucleic acid sequence encoding a polypeptide; A system for producing eukaryotic translatable mRNA, wherein transcription of said eukaryotic translatable mRNA is under the control of said prokaryotic promoter. 2. The true eukaryotic translatable mRNA of claim 1, wherein the eukaryotic translatable mRNA comprises a nucleic acid sequence encoding an element that promotes covalently closed circle formation after RNA transcription, an IRES, and a nucleic acid sequence encoding a polypeptide. A system for producing karyotic translatable mRNA. 3. The system for producing eukaryotic translatable mRNA according to claim 2, wherein the element promoting covalently closed circle formation after transcription of the RNA is a PIE sequence. 4. The system for producing eukaryotic translatable mRNA of claim 3, wherein the PIE sequence comprises bacteriophage T4 phage intron and exon elements. For producing a eukaryotic translatable mRNA according to claim 1, wherein the eukaryotic specific element for translation into protein comprises a viral or non-viral eukaryotic internal ribosome entry site (IRES) element. system. The eukaryotic translation of claim 1, wherein the IRES is an IRES selected from the group consisting of a cricket paralysis virus (CrPV) IRES, a foot and mouth disease virus (FMDV) IRES, and a swine fever virus (CSFV) IRES. A system for producing possible mRNA. A system for producing eukaryotic translatable mRNA according to claim 1, wherein the bacterium is a non-pathogenic bacterium engineered to have at least one invasion factor. 8. The system for producing eukaryotic translatable mRNA according to claim 7, wherein the invasion factor is encoded by the inv or hlyA gene. 2. The system for producing eukaryotic translatable mRNA of claim 1 , wherein the expression cassette encoding the eukaryotic translatable mRNA is engineered to reside on the chromosome of the bacterium. A system for producing eukaryotic translatable mRNA according to claim 1 , wherein the expression cassette is carried on a plasmid comprising a sequence encoding at least one mRNA molecule containing eukaryotic translatable elements. . 2. A system for producing eukaryotic translatable mRNA according to claim 1 , wherein the expression cassette comprises a polyA sequence . 12. A system for producing eukaryotic translatable mRNA according to claim 11 , wherein the polyA sequence contains no more than 500 adenine nucleotides . A system for producing eukaryotic translatable mRNA according to any one of claims 1 to 12, wherein the mRNA encodes a viral polypeptide or a fragment thereof. Administration to human patients or animal subjects may be intramuscular, parenteral, intravenous, intradermal, subcutaneous, intracardiac, intracerebral, intravitreal, intranasal, rectal, oral, intravaginal, inhalation, intraperitoneal, or bone. A system for producing eukaryotic translatable mRNA in eukaryotic cells according to any one of claims 1 to 13, which may be selected from the group consisting of intra-, intra-oral and intra-urethral administration. including a bacterium engineered to have at least one expression cassette encoding a eukaryotic translatable mRNA comprising a eukaryotic promoter, an internal ribosome entry sequence (IRES), and a nucleic acid sequence encoding a polypeptide. , a system for producing eukaryotic translatable mRNA, wherein transcription of said eukaryotic translatable mRNA is under the control of said eukaryotic promoter. system.