Highly efficient genome editing by CRISPR-Cpf1 using CRISPR RNA with a uridinylate-rich 3′-overhang

被引：122

作者：

Moon, Su Bin ^{[1
,2
]}

Lee, Jeong Mi ^{[1
,2
]}

Kang, Jeong Gu ^{[1
]}

Lee, Nan-Ee ^{[1
,2
]}

Ha, Dae-In ^{[1
,2
]}

Kim, Do Yon ^{[1
,2
]}

Kim, Sun Hee ^{[1
]}

Yoo, Kwangsun ^{[1
,2
]}

Kim, Daesik ^{[3
]}

Ko, Jeong-Heon ^{[1
,2
]}

Kim, Yong-Sam ^{[1
,2
]}

机构：

[1] KRIBB, Genome Editing Res Ctr, 125 Gwahak Ro, Daejeon 34141, South Korea

[2] Korea Univ Sci & Technol UST, KRIBB Sch Biosci, Dept Biomol Sci, 217 Gajeong Ro, Daejeon 34113, South Korea

[3] Seoul Natl Univ, Dept Chem, Seoul 08826, South Korea

来源：

NATURE COMMUNICATIONS | 2018年 / 9卷

基金：

新加坡国家研究基金会;

关键词：

DNA CLEAVAGE; T-CELLS; CPF1; COMPLEX; GENE; RESISTANCE; NUCLEASES; CAS9; SPECIFICITIES; ENDONUCLEASE;

D O I：

10.1038/s41467-018-06129-w

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：

Genome editing has been harnessed through the development of CRISPR system, and the CRISPR from Prevotel la and Francisella 1 (Cpf1) system has emerged as a promising alternative to CRISPR-Cas9 for use in various circumstances. Despite the inherent multiple advantages of Cpf1 over Cas9, the adoption of Cpf1 has been unsatisfactory because of target-dependent insufficient indel efficiencies. Here, we report an engineered CRISPR RNA (crRNA) for highly efficient genome editing by Cpf1, which includes a 20-base target-complementary sequence and a uridinylate-rich 3'-overhang. When the crRNA is transcriptionally produced, crRNA with a 20-base target-complementary sequence plus a U(4)AU(4) 3'-overhang is the optimal configuration. U-rich crRNA also maximizes the utility of the AsCpf1 mutants and multiplexing genome editing using mRNA as the source of multiple crRNAs. Furthermore, U-rich crRNA enables a highly safe and specific genome editing using Cpf1 in human cells, contributing to the enhancement of a genome-editing toolbox.

引用

页数：11

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