Enhanced mammalian genome editing by new Cas12a orthologs with optimized crRNA scaffolds

Fei Teng; Jing Li; Tongtong Cui; Kai Xu; Lu Guo; Qingqin Gao; Guihai Feng; Chuanyuan Chen; Dali Han; Qi Zhou; Wei Li

doi:10.1186/s13059-019-1620-8

Enhanced mammalian genome editing by new Cas12a orthologs with optimized crRNA scaffolds

Genome Biol. 2019 Feb 5;20(1):15. doi: 10.1186/s13059-019-1620-8.

Authors

Fei Teng^{1

2

3}, Jing Li^{1

2

4}, Tongtong Cui^{1

2

3}, Kai Xu^{1

2

3}, Lu Guo^{1

2

3}, Qingqin Gao^{1

2

3}, Guihai Feng^{1

2}, Chuanyuan Chen^{3

5}, Dali Han^{3

5}, Qi Zhou^{6

7

8}, Wei Li^{9

10

11}

Affiliations

¹ State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, 100101, China.
² Institute for Stem Cell and Regeneration, Chinese Academy of Sciences, Beijing, 100101, China.
³ University of Chinese Academy of Sciences, Beijing, 100049, China.
⁴ School of Life Sciences, University of Science and Technology of China, Hefei, 230026, China.
⁵ Key Laboratory of Genomic and Precision Medicine, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, 100101, China.
⁶ State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, 100101, China. qzhou@ioz.ac.cn.
⁷ Institute for Stem Cell and Regeneration, Chinese Academy of Sciences, Beijing, 100101, China. qzhou@ioz.ac.cn.
⁸ University of Chinese Academy of Sciences, Beijing, 100049, China. qzhou@ioz.ac.cn.
⁹ State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, 100101, China. liwei@ioz.ac.cn.
¹⁰ Institute for Stem Cell and Regeneration, Chinese Academy of Sciences, Beijing, 100101, China. liwei@ioz.ac.cn.
¹¹ University of Chinese Academy of Sciences, Beijing, 100049, China. liwei@ioz.ac.cn.

Abstract

CRISPR-Cas12a/Cpf1, a single RNA-guided endonuclease system, provides a promising tool for genome engineering. However, only three Cas12a orthologs have been employed for mammalian genome editing, and the editing efficiency as well as targeting coverage still requires improvements. Here, we harness six novel Cas12a orthologs for genome editing in human and mouse cells, some of which utilize simple protospacer adjacent motifs (PAMs) that remarkably increase the targeting range in the genomes. Moreover, we identify optimized CRISPR RNA (crRNA) scaffolds that can increase the genome editing efficiency of Cas12a.

Keywords: CRISPR-Cas12a/Cpf1; Genome editing; Human cells; Mouse cells; crRNA.

Publication types

Evaluation Study
Research Support, Non-U.S. Gov't

MeSH terms

Animals
CRISPR-Associated Proteins*
CRISPR-Cas Systems*
Gene Editing / methods*
Humans
Mice

Substances

CRISPR-Associated Proteins