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1. WO2020000439 - METHOD FOR KNOCKING OUT MOUSE CD366 GENE USING CRISPR/CAS9 SYSTEM

Publication Number WO/2020/000439
Publication Date 02.01.2020
International Application No. PCT/CN2018/093843
International Filing Date 29.06.2018
IPC
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N
MICRO-ORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICRO-ORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15
Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09
Recombinant DNA-technology
11
DNA or RNA fragments; Modified forms thereof
113
Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N
MICRO-ORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICRO-ORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15
Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09
Recombinant DNA-technology
63
Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
79
Vectors or expression systems specially adapted for eukaryotic hosts
85
for animal cells
86
Viral vectors
C12N 15/113 (2010.01)
C12N 15/86 (2006.01)
CPC
C12N 15/113
C12N 15/86
Applicants
  • 深圳市博奥康生物科技有限公司 SHENZHEN BIOCAN TECHNOLOGIES CO., LTD. [CN/CN]; 中国广东省深圳市 南山区西丽街道珠光路134号珠光创新科技园1栋509室 毛吉炎 MAO Jiyan Room 509, Building 1, Zhuguang Innovation Park No.134 Zhuguang Road, Xilii Street, Nanshan District Shenzhen, Guangdong 518055, CN
Inventors
  • 毛吉炎 MAO, Jiyan; CN
Priority Data
Publication Language Chinese (ZH)
Filing Language Chinese (ZH)
Designated States
Title
(EN) METHOD FOR KNOCKING OUT MOUSE CD366 GENE USING CRISPR/CAS9 SYSTEM
(FR) MÉTHODE D'INACTIVATION DU GÈNE CD366 DE SOURIS À L'AIDE D'UN SYSTÈME CRISPR/CAS9
(ZH) 一种应用CRISPR/Cas9系统对小鼠CD366基因进行敲除的方法
Abstract
(EN)
A method for knocking out a mouse CD366 gene using a CRISPR/Cas9 system: first acquiring a DNA sequence for a gRNA identification region of a mouse CD366 gene; next designing a gRNA that targets the DNA sequence, and constructing a gRNA in-vitro transcription vector containing a T7 promoter for the CD399 gene; and finally using the Cas9 and gRNA in-vitro transcription vector to acquire Cas9mRNA and gRNA, the Cas9mRNA and gRNA being acquired by means of injecting fertilised eggs.
(FR)
L'invention concerne une méthode d'inactivation d'un gène CD366 de souris à l'aide d'un système CRISPR/Cas9 comprenant les étapes suivantes : tout d'abord l'acquisition d'une séquence d'ADN pour une région d'identification d'ARNg d'un gène CD366 de souris ; puis la conception d'un ARNg qui cible la séquence d'ADN, et la construction d'un vecteur de transcription in vitro d'ARNg contenant un promoteur T7 pour le gène CD399 ; et enfin l'utilisation du vecteur de transcription d'ARNg et Cas9 in vitro pour l'acquisition de Cas9mRNA et de l'ARNg, le Cas9mRNA et l'ARNg étant acquis par injection d’œufs fécondés.
(ZH)
一种应用CRISPR/Cas9系统对小鼠CD366基因进行敲除的方法,其是首先获得针对小鼠CD366基因的gRNA识别区的DNA序列;其次设计靶向DNA序列的gRNA,构建针对CD366基因且含T7启动子的gRNA体外转录载体;最后利用Cas9和gRNA的体外转录载体获得Cas9mRNA、gRNA,通过受精卵注射获得的Cas9mRNA和gRNA。
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