Efficient transcriptional gene repression by type V-A CRISPR-Cpf1 from Eubacterium eligens = 유박테리움 eligens 유래 유전자가위를 이용한 효율적 유전자 발현 조절

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Title
Efficient transcriptional gene repression by type V-A CRISPR-Cpf1 from Eubacterium eligens = 유박테리움 eligens 유래 유전자가위를 이용한 효율적 유전자 발현 조절
Author(s)
Seong Keun KimHaseong Kim; Woo-Chan Ahn; Kwang Hyun ParkEui-Jeon WooDae-Hee LeeSeung Goo Lee
Bibliographic Citation
ACS Synthetic Biology, vol. 6, pp. 1273-1282
Publication Year
2017
Abstract
Clustered regularly interspaced short palindromic repeats interference (CRISPRi) is an emerging technology for artificial gene regulation. Type II CRISPR-Cas endonuclease Cas9 is the most widely used protein for gene regulation with CRISPRi. Here, we present type V-A CRISPR-Cas endonuclease Cpf1-based CRISPRi. We constructed an l-rhamnose-inducible CRISPRi system with DNase-deactivated Cpf1 from Eubacterium eligens (EedCpf1) and compared its performance with catalytically deactivated Cas9 from Streptococcus pyogenes (SpdCas9). In contrast to SpdCas9, EedCpf1 showed stronger gene repression when it was targeted to the template strand than when it was targeted to the nontemplate strand of the 5′ untranslated region or coding DNA sequences. EedCpf1 exhibited no strand bias when targeted to the promoter, and preferentially used the 5′-TTTV-3′ (V = A, G, or C) protospacer adjacent motif. Multiplex repression of the EedCpf1-based CRISPRi system was demonstrated using episomal and chromosomal gene targets. Our findings will guide an efficient EedCpf1-mediated CRISPRi genetic control
Keyword
CRISPRideactivated Cas9deactivated Cpf1Eubacterium eligensprotospacer adjacent motifStreptococcus pyogenes
ISSN
2161-5063
Publisher
Amer Chem Soc
DOI
http://dx.doi.org/10.1021/acssynbio.6b00368
Type
Article
Appears in Collections:
Synthetic Biology and Bioengineering Research Institute > Synthetic Biology Research Center > 1. Journal Articles
Critical Diseases Diagnostics Convergence Research Center > 1. Journal Articles
Division of Biomedical Research > Disease Target Structure Research Center > 1. Journal Articles
Synthetic Biology and Bioengineering Research Institute > 1. Journal Articles
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