Efficient Transcriptional Gene Repression by Type V-A CRISPR-Cpf1 from Eubacterium eligens

• DOI: 10.1021/acssynbio.6b00368
• Publication Date: 2017-04-05T08:01:23Z
• Authors: Seong Keun Kim; Haseong Kim; Woo-Chan Ahn; Kwang-Hyun Park; Eui-Jeon Woo; Dae-Hee Lee; Seung-Goo Lee
• Keywords: Eubacterium

Clustered regularly interspaced short palindromic repeats interference (CRISPRi) is an emerging technology for artificial gene regulation. Type II CRISPR-Cas endonuclease Cas9 the most widely used protein regulation with CRISPRi. Here, we present type V-A Cpf1-based We constructed l-rhamnose-inducible CRISPRi system DNase-deactivated Cpf1 from Eubacterium eligens (EedCpf1) and compared its performance catalytically deactivated Streptococcus pyogenes (SpdCas9). In contrast to SpdCas9, EedCpf1 showed stronger repression when it was targeted template strand than nontemplate of 5' untranslated region or coding DNA sequences. exhibited no bias promoter, preferentially 5'-TTTV-3' (V = A, G, C) protospacer adjacent motif. Multiplex EedCpf1-based demonstrated using episomal chromosomal targets. Our findings will guide efficient EedCpf1-mediated genetic control.