A modified glycosylase base editor without predictable <scp>DNA</scp> off‐target effects-Reference-Cited by-同舟云学术

A modified glycosylase base editor without predictable DNA off‐target effects

Published:2024-06-30 Issue: Volume: Page:
ISSN:0014-5793
Container-title:FEBS Letters
language:en
Short-container-title:FEBS Letters

Author:

Lian Meng¹²,Chen Tao³,Chen Min³,Peng Xiaohua³,Yang Yang⁴,Luo Xian³,Chi Yue³,Wang Jinling³,Tang Chengcheng³,Zhou Xiaoqing³,Zhang Kun³,Qin Chuan¹,Lai Liangxue¹²³⁴,Zhou Jizeng⁴,Zou Qingjian³^ORCID

Affiliation:

1. Institute of Laboratory Animal Sciences Chinese Academy of Medical Sciences & Peking Union Medical College Beijing China

2. Research Unit of Generation of Large Animal Disease Models Guangzhou China

3. Guangdong Provincial Key Laboratory of Large Animal Models for Biomedicine, South China Institute of Large Animal Models for Biomedicine, School of Biotechnology and Health Sciences Wuyi University Jiangmen China

4. CAS Key Laboratory of Regenerative Biology, Guangzhou Institutes of Biomedicine and Health Chinese Academy of Sciences Guangzhou China

Abstract

Glycosylase base editor (GBE) can induce C‐to‐G transversion in mammalian cells, showing great promise for the treatment of human genetic disorders. However, the limited efficiency of transversion and the possibility of off‐target effects caused by Cas9 restrict its potential clinical applications. In our recent study, we have successfully developed TaC9‐CBE and TaC9‐ABE by separating nCas9 and deaminase, which eliminates the Cas9‐dependent DNA off‐target effects without compromising editing efficiency. We developed a novel GBE called TaC9‐GBEYE1, which utilizes the deaminase and UNG‐nCas9 guided by TALE and sgRNA, respectively. TaC9‐GBEYE1 showed comparable levels of on‐target editing efficiency to traditional GBE at 19 target sites, without any off‐target effects caused by Cas9 or TALE. The TaC9‐GBEYE1 is a safe tool for gene therapy.

Funder

National Natural Science Foundation of China

Natural Science Foundation of Guangdong Province

National Key Research and Development Program of China Stem Cell and Translational Research

Publisher

Wiley

Link

https://febs.onlinelibrary.wiley.com/doi/pdf/10.1002/1873-3468.14970

Reference29 articles.

1. CRISPR C-to-G base editors for inducing targeted DNA transversions in human cells

2. Programmable editing of a target base in genomic DNA without double-stranded DNA cleavage

3. Base Excision Repair

4. Glycosylase base editors enable C-to-A and C-to-G base changes

5. AGBE: a dual deaminase-mediated base editor by fusing CGBE with ABE for creating a saturated mutant population with multiple editing patterns