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1. WO2006122443 - THE METHOD FOR KNOCKING-OUT THE FISH GENE

Publication Number WO/2006/122443
Publication Date 23.11.2006
International Application No. PCT/CN2005/000677
International Filing Date 16.05.2005
IPC
C12N 15/63 2006.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
C12N 15/11 2006.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
11DNA or RNA fragments; Modified forms thereof
C12N 15/12 2006.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
11DNA or RNA fragments; Modified forms thereof
12Genes encoding animal proteins
C12N 15/89 2006.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
87Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
89using microinjection
A01K 67/027 2006.01
AHUMAN NECESSITIES
01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
KANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
67Rearing or breeding animals, not otherwise provided for; New breeds of animals
027New breeds of vertebrates
CPC
A01K 2217/075
AHUMAN NECESSITIES
01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
KANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
2217Genetically modified animals
07Animals genetically altered by homologous recombination
075inducing loss of function, i.e. knock out
A01K 2227/40
AHUMAN NECESSITIES
01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
KANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
2227Animals characterised by species
40Fish
A01K 2267/02
AHUMAN NECESSITIES
01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
KANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
2267Animals characterised by purpose
02Animal zootechnically ameliorated
A01K 67/0276
AHUMAN NECESSITIES
01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
KANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
67Rearing or breeding animals, not otherwise provided for; New breeds of animals
027New breeds of vertebrates
0275Genetically modified vertebrates, e.g. transgenic
0276Knockout animals
C12N 15/8509
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
79Vectors or expression systems specially adapted for eukaryotic hosts
85for animal cells
8509for producing genetically modified animals, e.g. transgenic
Applicants
  • 中山大学 ZHONGSHAN UNIVERSITY [CN]/[CN] (AllExceptUS)
  • 中国科学院水生生物研究所 INSTITUTE OF HYDROBIOLOGY, CHINESE ACADEMY OF SCIENCES [CN]/[CN] (AllExceptUS)
  • 吴玉萍 WU, Yuping [CN]/[CN] (UsOnly)
  • 朱作言 ZHU, Zuoyan [CN]/[CN] (UsOnly)
  • 徐安龙 XU, Anlong [CN]/[CN] (UsOnly)
  • 张广献 ZHANG, Guangxian [CN]/[CN] (UsOnly)
  • 胡炜 HU, Wei [CN]/[CN] (UsOnly)
  • 熊茜 XIONG, Qian [CN]/[CN] (UsOnly)
  • 罗芳 LUO, Fang [CN]/[CN] (UsOnly)
  • 崔彩媚 CUI, Caimei [CN]/[CN] (UsOnly)
Inventors
  • 吴玉萍 WU, Yuping
  • 朱作言 ZHU, Zuoyan
  • 徐安龙 XU, Anlong
  • 张广献 ZHANG, Guangxian
  • 胡炜 HU, Wei
  • 熊茜 XIONG, Qian
  • 罗芳 LUO, Fang
  • 崔彩媚 CUI, Caimei
Agents
  • 广州新诺专利商标事务所有限公司 GUANGZHOU SINO PATENT & TRADEMARK AGENCY CO., LTD.
Priority Data
Publication Language Chinese (ZH)
Filing Language Chinese (ZH)
Designated States
Title
(EN) THE METHOD FOR KNOCKING-OUT THE FISH GENE
(FR) PROCEDE D'ELIMINATION DU GENE DES POISSONS
(ZH) 鱼类基因剔除的方法
Abstract
(EN)
The present invention disclosed a method for knocking-out the fish gene, comprising the steps of: 1) isolating the fish's substituting gene sequence, target gene sequence and flaking sequence thereof; 2) providing the target gene sequence of fish as the target DNA fragment for the homologous recombination; 3) constructing the homologous recombination vectors; 4) introducing the homologous recombination vectors into the fish oosperms by using micro-injections after artificial fertilization and before the first cleavage, and then incubating; 5) selecting the embryo carrying the markers with positive direction only to obtain the chimera with homologous recombination; 6) cross breeding the chimera to obtain homozygote. The method would avoid the random integration of the exogenous gene, and eliminate people's suspicion about the safety for transgenic organism, due to the fish gene knock-out by using homologous recombination. The present invention also established effective gene homologous recombination technology by constructing homologous recombination vectors, and made it possible to introduce site-directed and quantified mutagenesis into the genetic structure of fish as designed, thereby to modify directionally the genetic structure and properties of the cell and the organism per se.
(FR)
L'invention concerne un procédé d'élimination du gène des poissons comprenant les étapes: 1) isolation de la séquence génétique de substitution du poisson, de la séquence génétique cible et la séquence d'arrachage; 2) apport de la séquence génétique cible du poisson comme fragment ADN cible pour la recombinaison homologue; 3) construction des vecteurs de recombinaison homologues; 4) introduction des vecteurs de recombinaison homologues dans le sperme du poisson en utilisant des microinjections après fécondation artificielle et avant la premier clivage, suivie de l'incubation; 5) sélection de l'embryon portant les marqueurs à direction positives afin d'obtenir uniquement le chimère à recombinaison analogue; 6) croisement entre races des chimères afin d'obtenir un homozygote. Le procédé permet d'éviter l'intégration aléatoire du gène exogène et de rassurer les gens quant à son innocuité pour l'organisme transgénique en raison de l'élimination du gène des poissons par utilisation de la recombinaison homologue. L'invention concerne également la technologie efficace de recombinaison homologue génétique par construction de vecteurs de recombinaisons homologues et rend possible l'introduction d'une mutagenèse orientée sur le site et quantifiée dans la structure génétique du poisson tel quel, avec pour effet de modifier directionnellement la structure génétique et les propriétés de la cellule et de l'organisme à proprement parlé.
Also published as
EP5749704
Latest bibliographic data on file with the International Bureau