Engineering the Direct Repeat Sequence of CrRNA for Optimization of FnCpf1-Mediated Genome Editing in Human Cells

Overview

Journal Mol Ther

Publisher Cell Press

Specialties Molecular Biology
Pharmacology

Date 2018 Oct 3

PMID 30274789

Citations 13

Authors

Li Lin

Xiubin He

Tianyuan Zhao

Lingkai Gu

Yeqing Liu

Xiaoyu Liu

Hongyan Liu

Fayu Yang

Mengjun Tu

Lianchao Tang

Xianglian Ge

Changbao Liu

Junzhao Zhao

Zongming Song

Jia Qu

Feng Gu

Affiliations

Soon will be listed here.

Abstract

FnCpf1-mediated genome-editing technologies have enabled a broad range of research and medical applications. Recently, we reported that FnCpf1 possesses activity in human cells and recognizes a more compatible PAM (protospacer adjacent motif, 5'-KYTV-3'), compared with the other two commonly used Cpf1 enzymes (AsCpf1 and LbCpf1), which requires a 5'-TTTN-3' PAM. However, due to the efficiency and fidelity, FnCpf1-based clinical and basic applications remain a challenge. The direct repeat (DR) sequence is one of the key elements for FnCpf1-mediated genome editing. In principle, its engineering should influence the corresponding genome-editing activity and fidelity. Here we showed that the DR mutants [G(-9)A and U(-7)A] could modulate FnCpf1 performance in human cells, enabling enhancement of both genome-editing efficiency and fidelity. These newly identified features will facilitate the design and optimization of CRISPR-Cpf1-based genome-editing strategies.

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