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1. WO2020000440 - METHOD FOR TARGETED KNOCKOUT OF HUMAN ASP2 GENE BASED ON CRISPR/CAS9 AND SPECIFIC GRNA THEREOF

Publication Number WO/2020/000440
Publication Date 02.01.2020
International Application No. PCT/CN2018/093844
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
C12N 15/113 (2010.01)
CPC
C12N 15/113
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 TARGETED KNOCKOUT OF HUMAN ASP2 GENE BASED ON CRISPR/CAS9 AND SPECIFIC GRNA THEREOF
(FR) PROCÉDÉ D'INACTIVATION CIBLÉE DU GÈNE ASP2 HUMAIN FONDÉ SUR CRISPR/CAS9 ET ARNG SPÉCIFIQUE ASSOCIÉ
(ZH) CRISPR/Cas9靶向敲除人ASP2基因的方法及其特异性gRNA
Abstract
(EN)
Provided are a method for a targeted knockout of human ASP2 gene based on CRISPR/Cas9 and a specific gRNA thereof. The method comprises: according to design principles of CRISPR/Cas9, designing two target sites upstream and downstream of human ASP2 gene, respectively, synthesizing corresponding oligonucleotide sequences, ligating into px459 vector to construct a recombinant plasmid, transfecting the recombinant plasmid into a human esophageal cancer cell line, and performing specific knockout of human ASP2 gene.
(FR)
L'invention concerne un procédé pour une inactivation ciblée du gène ASP2 humain fondé sur CRISPR/Cas9 et un ARNg spécifique associé. Le procédé comprend les étapes suivantes : selon des principes de conception de CRISPR/Cas9, la conception de deux sites cibles en amont et en aval du gène ASP2 humain, respectivement, la synthèse de séquences oligonucléotidiques correspondantes, la ligature dans un vecteur px459 pour construire un plasmide recombinant, la transfection du plasmide recombinant dans une lignée cellulaire de cancer de l'œsophage humain, et la réalisation d'une inactivation spécifique du gène ASP2 humain.
(ZH)
提供一种CRISPR/Cas9靶向敲除人ASP2基因的方法及其特异性gRNA,所述方法包括:根据CRISPR/Cas9的设计原则,在人ASP2基因的上下游设计两个靶位点,合成相应的寡核苷酸序列,连接至px459载体上构建重组质粒,将重组质粒转染人食管癌细胞株,特异性敲除人ASP2基因。
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