Multiplex gene regulation by CRISPR-ddCpf1

doi:10.1038/celldisc.2017.18

	Multiplex gene regulation by CRISPR-ddCpf1
	Zhang, Xiaochun1,2,3 ; Wang, Jingman 4,5; Cheng, Qiuxiang 6; Zheng, Xuan 1; Zhao, Guoping 1; Wang, Jin 1
	2017-06-06
发表期刊	CELL DISCOVERY
ISSN	2056-5968
卷号	3
发表状态	已发表
DOI	10.1038/celldisc.2017.18
摘要	The clustered regularly interspaced short palindromic repeats (CRISPR)/dCas9 system has been widely applied in both transcriptional regulation and epigenetic studies. However, for multiple targets, independent expression of multiple single guide RNAs (sgRNAs) is needed, which is less convenient. To address the problem, we employed a DNase-dead Cpf1 mutant (ddCpf1) for multiplex gene regulation. We demonstrated that ddCpf1 alone could be employed for gene repression in Escherichia coli, and the repression was more effective with CRISPR RNAs (crRNAs) specifically targeting to the template strand of its target genes, which was different from that of dCas9. When targeting the promoter region, both strands showed effective repression by the ddCpf1/crRNA complex. The whole-transcriptome RNA-seq technique was further employed to demonstrate the high specificity of ddCpf1-mediated repression. Besides, we proved that the remaining RNase activity in ddCpf1 was capable of processing a precursor CRISPR array to simply generate multiple mature crRNAs in vivo, facilitating multiplex gene regulation. With the employment of this multiplex gene regulation strategy, we also showed how to quickly screen a library of candidate targets, that is, the two-component systems in E. coli. Therefore, based on our findings here, the CRISPR-ddCpf1 system may be further developed and widely applied in both biological research and clinical studies.
关键词	CRISPR Cpf1 DNase-dead Cpf1 (ddCpf1) CRISPRi multiplex gene regulation
收录类别	SCI
语种	英语
资助项目	National Natural Science Foundation of China[31430004] ; National Natural Science Foundation of China[31421061]
WOS研究方向	Cell Biology
WOS类目	Cell Biology
WOS记录号	WOS:000414915700001
出版者	NATURE PUBLISHING GROUP
WOS关键词	CRISPR-CAS SYSTEMS ; ESCHERICHIA-COLI ; RNA-SEQ ; TARGETED MUTAGENESIS ; 2-COMPONENT SYSTEMS ; HUMAN-CELLS ; CPF1 ; REPRESSION ; TRANSCRIPTION ; ACTIVATION
原始文献类型	Article
引用统计
文献类型	期刊论文
条目标识符	https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/10055
专题	生命科学与技术学院生命科学与技术学院_硕士生
通讯作者	Wang, Jin
作者单位	1.Chinese Acad Sci, Key Lab Synthet Biol, Inst Plant Physiol & Ecol, Shanghai Inst Biol Sci, Shanghai, Peoples R China 2.Shanghai Tech Univ, Sch Life Sci & Technol, Shanghai, Peoples R China 3.Univ Chinese Acad Sci, Beijing, Peoples R China 4.Shenzhen Univ, State Engn Lab Med Key Technol Applicat Synthet, Shenzhen Peoples Hosp 2, Affiliated Hosp 1, Shenzhen, Peoples R China 5.Sun Yat Sen Univ, Canc Ctr, Guangzhou, Guangdong, Peoples R China 6.Shanghai Tolo Biotechnol Co Ltd, Shanghai, Peoples R China
第一作者单位	生命科学与技术学院
推荐引用方式 GB/T 7714	Zhang, Xiaochun,Wang, Jingman,Cheng, Qiuxiang,et al. Multiplex gene regulation by CRISPR-ddCpf1[J]. CELL DISCOVERY,2017,3.
APA	Zhang, Xiaochun,Wang, Jingman,Cheng, Qiuxiang,Zheng, Xuan,Zhao, Guoping,&Wang, Jin.(2017).Multiplex gene regulation by CRISPR-ddCpf1.CELL DISCOVERY,3.
MLA	Zhang, Xiaochun,et al."Multiplex gene regulation by CRISPR-ddCpf1".CELL DISCOVERY 3(2017).