Efficient multinucleotide deletions using deaminase-Cas9 fusions in human cells

Siyu Chen; Zhiquan Liu; Hao Yu; Liangxue Lai; Zhanjun Li

doi:10.1016/j.jgg.2022.03.007

Volume 49 Issue 10

Oct. 2022

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Article Navigation > Journal of Genetics and Genomics > 2022 > 49(10): 927-933

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Efficient multinucleotide deletions using deaminase-Cas9 fusions in human cells

doi: 10.1016/j.jgg.2022.03.007

a. Key Laboratory of Zoonosis Research, Ministry of Education, College of Animal Science, Jilin University, Changchun, Jilin 130062, China;
b. CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, Guangdong 510530, China;
c. Guangzhou Regenerative Medicine and Health Guang Dong Laboratory (GRMH-GDL), Guangzhou, Guangdong 510005, China;
d. Institute for Stem Cell and Regeneration, Chinese Academy of Sciences, Beijing 100101, China

Funds:

Development Program of Guangzhou Regenerative Medicine and Health Guangdong Laboratory (2018GZR110104004).

The authors thank Peiran Hu and Nannan Li at the Embryo Engineering Center for critical technical assistance. This work was supported by the National Key Research and Development Program of China Stem Cell and Translational Research (2019YFA0110700), the Program for Changjiang Scholars and Innovative Research Team in University (No.IRT_16R32), the Strategic Priority Research Program of the Chinese Academy of Sciences (XDA16030501, XDA16030503), and Key Research &

Received Date: 2021-10-20
Accepted Date: 2022-03-22
Rev Recd Date: 2022-03-06
Publish Date: 2022-04-11

Abstract

Abstract

CRISPR/Cas9 system is a robust genome editing platform in biotechnology and medicine. However, it generally produces small insertions/deletions (indels, typically 1–3 bp) but rarely induces larger deletions in specific target sites. Here, we report a cytidine deaminase-Cas9 fusion-induced deletion system (C-DEL) and an adenine deaminase-Cas9 fusion-induced deletion system (A-DEL) by combining Cas9 with rat APOBEC1 (rA1) and TadA 8e, respectively. Both C-DEL and A-DEL improve the efficiency of deletions compared with the conventional Cas9 system in human cells. In addition, the C-DEL system generates a considerable fraction of predictable multinucleotide deletions from 5′-deaminated C bases to the Cas9-cleavage site and increases the proportion of larger deletions at the target loci. Taken together, the C-DEL and A-DEL systems provide a practical strategy for producing efficient multinucleotide deletions, expanding the CRISPR/Cas9 toolsets for gene modifications in human cells.
- CRISPR/Cas9,
- Deaminase,
- C-DEL,
- A-DEL

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References(34)

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