基本信息:
- 专利标题： 利用CRISPR/Cas9基因编辑系统敲除小菜蛾APN1基因及其应用
- 专利标题（英）：Method for knocking out plutella xylostella APN1 gene with CRISPR/Cas9 gene editing system and application of method
- 申请号：CN201811642687.3 申请日：2018-12-29
- 公开(公告)号：CN109593762A 公开(公告)日：2019-04-09
- 发明人： 张友军 , 郭兆将 , 孙丹 , 康师 , 郭乐 , 周君雷 , 龚莉君 , 覃舰莹 , 朱流红 , 白杨 , 罗亮
- 申请人： 中国农业科学院蔬菜花卉研究所
- 申请人地址： 北京市海淀区中关村南大街12号
- 专利权人： 中国农业科学院蔬菜花卉研究所
- 当前专利权人： 中国农业科学院蔬菜花卉研究所
- 当前专利权人地址： 北京市海淀区中关村南大街12号
- 代理机构： 北京知本村知识产权代理事务所
- 代理人： 刘江良
- 主分类号： C12N15/113
- IPC分类号： C12N15/113 ; C12N15/90
The invention discloses a method for knocking out a non-model organism plutella xylostella APN1 gene with CRISPR/Cas9 and an application of the method in molecular research on Bt Cry1Ac insecticidal protein resistance mechanisms. According to the method, a specific sgRNA target sequence of the plutella xylostella APN1 gene is designed and synthesized, sgRNA and Cas9 protein are mixed, the mixtureis microscopically injected into the posterior pole part of early embryonic disc stage eggs of plutella xylostella, plutella xylostella APN1 gene homozygous mutant populations with stable heredity arescreened out by a CRISPR/Cas9 gene editing system and applied to toxicity bioassay of Bt Cry1Ac insecticidal protein, which proves that plutella xylostella APN1 gene mutation can cause high resistance to Bt Cry1Ac insecticidal protein. The method is simple and efficient, time and labor are saved, technical support is provided for deep research on functional genes in non-model organisms, and a foundation is laid for exploring new pest control strategies.