DK2155880T3 - Modificering af glycoproteinproduktion i planter - Google Patents

Modificering af glycoproteinproduktion i planter Download PDF

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Publication number
DK2155880T3
DK2155880T3 DK08772802.8T DK08772802T DK2155880T3 DK 2155880 T3 DK2155880 T3 DK 2155880T3 DK 08772802 T DK08772802 T DK 08772802T DK 2155880 T3 DK2155880 T3 DK 2155880T3
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Prior art keywords
plant
protein
interest
nucleotide sequence
seq
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DK08772802.8T
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English (en)
Inventor
Marc-Andre D'aoust
Estelle Marquet-Blouin
Carole Burel
Loic Faye
Patrice Lerouge
Véronique Gomord
Stéphanie Aquin
Christophe Rihouey
Thomas Paccalet
Christophe Sourrouille
Muriel Bardor
Louis-Philippe Vezina
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Medicago Inc
Centre Nat Rech Scient
Univ Rouen
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Publication of DK2155880T3 publication Critical patent/DK2155880T3/da

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    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8242—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits
    • C12N15/8257—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits for the production of primary gene products, e.g. pharmaceutical products, interferon
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    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8216—Methods for controlling, regulating or enhancing expression of transgenes in plant cells
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    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8216—Methods for controlling, regulating or enhancing expression of transgenes in plant cells
    • C12N15/8218—Antisense, co-suppression, viral induced gene silencing [VIGS], post-transcriptional induced gene silencing [PTGS]
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8242—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits
    • C12N15/8243—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine
    • C12N15/8245—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine involving modified carbohydrate or sugar alcohol metabolism, e.g. starch biosynthesis
    • C—CHEMISTRY; METALLURGY
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    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8242—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits
    • C12N15/8257—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits for the production of primary gene products, e.g. pharmaceutical products, interferon
    • C12N15/8258—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits for the production of primary gene products, e.g. pharmaceutical products, interferon for the production of oral vaccines (antigens) or immunoglobulins
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    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/10—Transferases (2.)
    • C12N9/1048—Glycosyltransferases (2.4)
    • C12N9/1051—Hexosyltransferases (2.4.1)

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  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
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  • Engineering & Computer Science (AREA)
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  • Wood Science & Technology (AREA)
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  • Plant Pathology (AREA)
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  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
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  • Immunology (AREA)
  • Virology (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Peptides Or Proteins (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
  • Cultivation Of Plants (AREA)

Claims (16)

  1. MODIFICERING AF GLYCOPROTEINPRODUKTION I PLANTER Patentkrav
    1. Fremgangsmåde til syntetisering af et protein af interesse med reduceret fucosylation og xylosylation omfattende indføring i en plant, en del af en plante eller en plantecelle, af en nukleotidsekvens omfattende fra 80 % til 100 % identitet med en nukleotidsekvens defineret ved SEQ ID NO: 17 og kodende for et hybridprotein omfattende en cytoplasmatisk hale, et transmembrandomæne, en stem-region (CTS domæne) af N-acetylglucosaminyltransferase (GNT1) kondenseret til et katalytisk domæne på beta-l,4-galactosyltransferase (GalT), hvilken første nukleotidsekvens er operationelt koblet med en første regulerende region, der er aktiv i planten, og en anden nukleotidsekvens til kodning for proteinet af interesse, hvilken anden nukleotidsekvens er operationelt koblet med en anden regulerende region, der er aktiv i planten, og transitorisk samtidig eksprimerer den første og anden nukleotidsekvens for at syntetisere proteinet af interesse omfattende mindre end 10 % fucosylation og xylosylation sammenlignet med samme protein af interesse produceret i en vildtype-plante.
  2. 2. Fremgangsmåde ifølge krav 1, hvori den første regulerende region er en plastocyanin-promoter eller en 35S-promoter, og den anden regulerende region er en plastocyanin-promoter eller en 35S-promoter.
  3. 3. Fremgangsmåde ifølge krav 1, hvori en tredje nukleotidsekvens eksprimeres i planten, hvilken tredje nukleotidsekvens koder for en silencing-suppressor, hvilken tredje nukleotidsekvens er operationelt koblet med en tredje regulerende region, der er aktiv i planten.
  4. 4. Fremgangsmåde ifølge krav 3, hvor proteinet af interesse syntetiseres i en mængde på op til 1,5 g pr. kilogram plante-vådvægt.
  5. 5. Fremgangsmåde ifølge krav 3, hvori den tredje regulerende region er en plastocyanin-promoter eller en 35S-promoter.
  6. 6. Fremgangsmåde ifølge krav 2, hvori en tredje nukleotidsekvens eksprimeres i planten, hvilken tredje nukleotidsekvens koder for en silencing-suppressor, hvilken tredje nukleotidsekvens er operationelt koblet med en tredje regulerende region, der er aktiv i planten.
  7. 7. Fremgangsmåde ifølge krav 6, hvori den tredje regulerende region er en plastocyanin-promoter eller en 35S-promoter.
  8. 8. Fremgangsmåde ifølge krav 1, hvori proteinet af interesse er et antistof, et antigen eller en vaccine.
  9. 9. Fremgangsmåde ifølge krav 8, hvori den anden nukleotidsekvens, som koder for proteinet af interesse, omfatter en nukleotidsekvens 2A, som er operationelt koblet med en regulerende region 2A, der er aktiv i planten, og en nukleotidsekvens 2B, som er operationelt koblet med en regulerende region 2B, der er aktiv i planten, og produktet, som 2A og 2B hver især koder for, kombineres for at danne et antistof.
  10. 10. Fremgangsmåde ifølge krav 9, hvori den regulerende region 2A er en plastocyanin-promoter og den regulerende region 2B er en plastocyanin-promoter.
  11. 11. Nukleotidsekvens omfattende nukleotid 1 til 1062 af SEQ ID NO:17 (GNTl-GalT), hvori nukleotidsekvensen koder for et protein, der modificerer glycosylation af et protein af interesse.
  12. 12. Hybridprotein GNTl-GalT omfattende et CTS-domæne af N-acetylglucosaminyltransferase kondenseret til et katalytisk domæne på beta-l,4-galactosyltransferase, hvilket hybridprotein omfatter aminosyre 1 til 354 af sekvens SEQ ID NO: 18.
  13. 13. Plante, plantecelle eller et frø omfattende nukleotidsekvensen ifølge krav 11.
  14. 14. Plante, plantecelle eller et frø omfattende hybridproteinet ifølge krav 12.
  15. 15. Fremgangsmåde ifølge krav 3, hvor proteinet af interesse er et antistof og syntetiseres i en mængde på op til 1,5 g pr. kilogram blad-vådvægt.
  16. 16. Fremgangsmåde ifølge krav 1, hvori det syntetiserede protein af interesse mangler fucosylation, xylosylation eller begge dele.
DK08772802.8T 2007-06-15 2008-06-13 Modificering af glycoproteinproduktion i planter DK2155880T3 (da)

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Application Number Priority Date Filing Date Title
US94434407P 2007-06-15 2007-06-15
PCT/CA2008/001139 WO2008151440A1 (en) 2007-06-15 2008-06-13 Modifying glycoprotein production in plants

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DK2155880T3 true DK2155880T3 (da) 2016-12-05

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US (1) US20110008837A1 (da)
EP (1) EP2155880B1 (da)
JP (1) JP5783720B2 (da)
KR (2) KR20130075787A (da)
CN (3) CN103382487A (da)
AU (1) AU2008261527C1 (da)
BR (1) BRPI0812538B1 (da)
CA (2) CA2700180C (da)
DK (1) DK2155880T3 (da)
EG (1) EG26600A (da)
ES (1) ES2624776T3 (da)
HU (1) HUE031698T2 (da)
IL (1) IL202568A (da)
MA (1) MA31516B1 (da)
MX (1) MX2009013665A (da)
NZ (2) NZ581944A (da)
PL (1) PL2155880T3 (da)
PT (1) PT2155880T (da)
RU (2) RU2499053C2 (da)
SG (1) SG185249A1 (da)
SI (1) SI2155880T1 (da)
TN (1) TN2009000512A1 (da)
WO (1) WO2008151440A1 (da)
ZA (1) ZA201000207B (da)

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RU2499053C2 (ru) 2013-11-20
EP2155880A1 (en) 2010-02-24
SG185249A1 (en) 2012-11-29
ES2624776T3 (es) 2017-07-17
AU2008261527C1 (en) 2014-09-04
PL2155880T3 (pl) 2017-06-30
EP2155880A4 (en) 2010-08-04
ZA201000207B (en) 2013-09-25
MX2009013665A (es) 2010-06-02
RU2010101024A (ru) 2011-07-20
US20110008837A1 (en) 2011-01-13
IL202568A (en) 2015-02-26
SI2155880T1 (sl) 2017-03-31
WO2008151440A8 (en) 2010-03-04
AU2008261527A8 (en) 2010-01-21
KR20100037602A (ko) 2010-04-09
KR20130075787A (ko) 2013-07-05
WO2008151440A1 (en) 2008-12-18
EP2155880B1 (en) 2016-08-10
NZ581944A (en) 2012-03-30
RU2013137208A (ru) 2015-02-20
AU2008261527A1 (en) 2008-12-18
NZ598417A (en) 2013-06-28
JP2010531136A (ja) 2010-09-24
CA2700180C (en) 2013-01-29
CN101918559A (zh) 2010-12-15
TN2009000512A1 (en) 2011-03-31
CN104962579A (zh) 2015-10-07
EG26600A (en) 2014-03-23
CA2700180A1 (en) 2008-12-18
IL202568A0 (en) 2011-08-01
KR101385050B1 (ko) 2014-05-13
BRPI0812538B1 (pt) 2021-03-16
CA2795379A1 (en) 2008-12-18
HUE031698T2 (en) 2017-07-28
CN103382487A (zh) 2013-11-06
WO2008151440A4 (en) 2009-02-26
BRPI0812538A2 (pt) 2014-09-30
AU2008261527B8 (en) 2014-04-24
PT2155880T (pt) 2016-11-18
AU2008261527B2 (en) 2014-04-10
JP5783720B2 (ja) 2015-09-24
MA31516B1 (fr) 2010-07-01

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