CRISPR/Cas9 mediated multiplex genome editing and heritable mutagenesis of BmKu70 in Bombyx mori

被引：123

作者：

Ma, Sanyuan ^{[1
]}

Chang, Jiasong ^{[1
]}

Wang, Xiaogang ^{[1
]}

Liu, Yuanyuan ^{[1
]}

Zhang, Jianduo ^{[1
]}

Lu, Wei ^{[1
]}

Gao, Jie ^{[1
]}

Shi, Run ^{[1
]}

Zhao, Ping ^{[1
]}

Xia, Qingyou ^{[1
]}

机构：

[1] Southwest Univ, State Key Lab Silkworm Genome Biol, Chongqing 400716, Peoples R China

来源：

SCIENTIFIC REPORTS | 2014年 / 4卷

关键词：

ONE-STEP GENERATION; TARGETED MUTAGENESIS; SILKWORM; GENE; KU70; TRANSLOCATION; SYSTEM; SITE;

D O I：

10.1038/srep04489

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

CRISPR/Cas9, a bacterial adaptive immune system derived genome-editing technique, has become to be one of the most compelling topics in biotechnology. Bombyx mori is an economically important insect and a model organism for studying lepidopteran and arthropod biology. Here we reported highly efficient and multiplex genome editing in B. mori cell line and heritable site-directed mutagenesis of Bmku70, which is required for NHEJ pathway and also related to antigen diversity, telomere length maintenance and subtelomeric gene silencing, using CRISPR/Cas9 system. We established a simple and practicable method and obtained several Bmku70 knockout B. mori lines, and showed that the frequency of HR was increased in embryos of the Bmku70 knockout B. mori. The mutant lines obtained in this study could be a candidate genetic resource for efficient knock-in and fundamental research of DNA repair in B. mori. We also provided a strategy and procedure to perform heritable genome editing of target genes with no significant phenotype effect.

引用

页数：6

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