A Convenient Cas9-based Conditional Knockout Strategy for Simultaneously Targeting Multiple Genes in Mouse

doi:10.1038/s41598-017-00654-2

	A Convenient Cas9-based Conditional Knockout Strategy for Simultaneously Targeting Multiple Genes in Mouse
	Chen, Jiang 1,2; Du, Yinan 4; He, Xueyan 1,2; Huang, Xingxu3 ; Shi, Yun S.1,2
	2017-03-31
发表期刊	SCIENTIFIC REPORTS (IF:3.8[JCR-2023],4.3[5-Year])
ISSN	2045-2322
卷号	7
发表状态	已发表
DOI	10.1038/s41598-017-00654-2
摘要	The most powerful way to probe protein function is to characterize the consequence of its deletion. Compared to conventional gene knockout (KO), conditional knockout (cKO) provides an advanced gene targeting strategy with which gene deletion can be performed in a spatially and temporally restricted manner. However, for most species that are amphiploid, the widely used Cre-flox conditional KO (cKO) system would need targeting loci in both alleles to be loxP flanked, which in practice, requires time and labor consuming breeding. This is considerably significant when one is dealing with multiple genes. CRISPR/Cas9 genome modulation system is advantaged in its capability in targeting multiple sites simultaneously. Here we propose a strategy that could achieve conditional KO of multiple genes in mouse with Cre recombinase dependent Cas9 expression. By transgenic construction of loxPstop- loxP (LSL) controlled Cas9 (LSL-Cas9) together with sgRNAs targeting EGFP, we showed that the fluorescence molecule could be eliminated in a Cre-dependent manner. We further verified the efficacy of this novel strategy to target multiple sites by deleting c-Maf and MafB simultaneously in macrophages specifically. Compared to the traditional Cre-flox cKO strategy, this sgRNAs-LSL-Cas9 cKO system is simpler and faster, and would make conditional manipulation of multiple genes feasible.
收录类别	SCI
语种	英语
资助项目	Natural Science Foundation of Jiangsu Province[BK20140018]
WOS研究方向	Science & Technology - Other Topics
WOS类目	Multidisciplinary Sciences
WOS记录号	WOS:000397879800005
出版者	NATURE PUBLISHING GROUP
WOS关键词	GENOME-EDITING SPECIFICITY ; HUMAN-CELLS ; MICE ; CAS9 ; MACROPHAGES ; RECOMBINATION ; CRISPR-CAS9 ; GENERATION ; PROTEIN ; CRE
原始文献类型	Article
引用统计	正在获取...
文献类型	期刊论文
条目标识符	https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/1449
专题	生命科学与技术学院_PI研究组_黄行许组
通讯作者	Huang, Xingxu; Shi, Yun S.
作者单位	1.Nanjing Univ, Model Anim Res Ctr, State Key Lab Pharmaceut Biotechnol, Nanjing 210061, Jiangsu, Peoples R China 2.Nanjing Univ, Model Anim Res Ctr, MOE Key Lab Model Anim Dis Study, Nanjing 210061, Jiangsu, Peoples R China 3.Shanghai Tech Univ, Sch Life Sci & Technol, 100 Haike Rd,Pudong New Area, Shanghai 201210, Peoples R China 4.Anhui Med Univ, Dept Microbiol & Parasitol, Hefei 230032, Peoples R China
通讯作者单位	生命科学与技术学院
推荐引用方式 GB/T 7714	Chen, Jiang,Du, Yinan,He, Xueyan,et al. A Convenient Cas9-based Conditional Knockout Strategy for Simultaneously Targeting Multiple Genes in Mouse[J]. SCIENTIFIC REPORTS,2017,7.
APA	Chen, Jiang,Du, Yinan,He, Xueyan,Huang, Xingxu,&Shi, Yun S..(2017).A Convenient Cas9-based Conditional Knockout Strategy for Simultaneously Targeting Multiple Genes in Mouse.SCIENTIFIC REPORTS,7.
MLA	Chen, Jiang,et al."A Convenient Cas9-based Conditional Knockout Strategy for Simultaneously Targeting Multiple Genes in Mouse".SCIENTIFIC REPORTS 7(2017).