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

Su Moon, Jeong Mi Lee, Jeong Gu Kang, Nan-Ee Lee, Dae-In Ha, Do Yon Kim, Sun Hee Kim, Kwangsun Yoo, Daesik Kim, Jeong-Heon Ko and Yong-Sam Kim ()
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Su Moon: Genome Editing Research Center, KRIBB
Jeong Mi Lee: Genome Editing Research Center, KRIBB
Jeong Gu Kang: Genome Editing Research Center, KRIBB
Nan-Ee Lee: Genome Editing Research Center, KRIBB
Dae-In Ha: Genome Editing Research Center, KRIBB
Do Yon Kim: Genome Editing Research Center, KRIBB
Sun Hee Kim: Genome Editing Research Center, KRIBB
Kwangsun Yoo: Genome Editing Research Center, KRIBB
Daesik Kim: Seoul National University
Jeong-Heon Ko: Genome Editing Research Center, KRIBB
Yong-Sam Kim: Genome Editing Research Center, KRIBB

Nature Communications, 2018, vol. 9, issue 1, 1-11

Abstract: Abstract Genome editing has been harnessed through the development of CRISPR system, and the CRISPR from Prevotella 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 U4AU4 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.

Date: 2018
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DOI: 10.1038/s41467-018-06129-w

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