Title:
NOVEL GENE TARGETING METHOD
Document Type and Number:
WIPO Patent Application WO/2016/173555
Kind Code:
A1
Abstract:
Provided are a novel two-step gene targeting method and a nucleotide construct for gene targeting. The method can improve the gene targeting efficiency and accurately identify a target gene knock-out mutant. The method of the present invention comprises: firstly, efficiently replacing a target gene in a genome with a targeting box by homologous recombination, the targeting box consisting of a target gene activity variant, a marker gene and site-specific recombination sites; and secondly, resecting the targeting box by recombinase, leaving a site-specific recombination site on the target gene to generate a target gene knock-out mutant, and removing a recombinase expression vector from the knock-out mutant by using a counter selection marker in the recombinase expression vector.
Inventors:
JIANG YOUWEI (CN)
LI YUNFEI (CN)
LI YUNFEI (CN)
Application Number:
PCT/CN2016/080785
Publication Date:
November 03, 2016
Filing Date:
April 29, 2016
Export Citation:
Assignee:
HANGZHOU GENEKINE BIOTECH CO LTD (CN)
International Classes:
C12N1/16; C12N15/11; C12P21/00
Foreign References:
| CN1844401A | 2006-10-11 | |||
| CN101440352A | 2009-05-27 |
Other References:
ZAO, CHEN.: "Enhancement of the Gene Targeting Efficiency of Non-Conventional Yeasts by Increasing Genetic Redundancy", PLOS ONE, vol. 8, no. 3, 7 March 2013 (2013-03-07), pages 1 - 9, XP055325893, ISSN: 1932-6203
See also references of EP 3290518A4
See also references of EP 3290518A4
Attorney, Agent or Firm:
XU & PARTNERS, LLC. (CN)
上海一平知识产权代理有限公司 (CN)
上海一平知识产权代理有限公司 (CN)
Download PDF: