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Efficient replacement of long DNA fragments via non-homologous end joining at non-coding regions
Shanye Gu1,2,†,* , Jia Li3,† , Siyuan Li1,† , Jianbin Cao4 , Jiwen Bu3 , Yonggang Ren3 , Wenjie Du3 , Zhecong Chen1 , Chufan Xu1 , Mingcang Wang4 , Lai Jiang1,* , Cheng Huang2,* , Jiulin Du3,5,6,*
1Department of Anesthesiology and Surgical Intensive Care Unit, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China
2Drug Discovery Laboratory, School of Pharmacy, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
3Institute of Neuroscience, State Key Laboratory of Neuroscience, Center for Excellence in Brain Science and Intelligence Technology, Shanghai Research Center for Brain Science and Brain- Inspired Intelligence, Chinese Academy of Sciences, Shanghai 200031, China
4Department of Anesthesiology, Affiliated Taizhou Hospital, Wenzhou Medical University, Linhai 317000, China
5University of Chinese Academy of Sciences, Beijing 100049, China
6ShanghaiTech University, Shanghai 200031, China
†These authors contributed equally to this work.
*Correspondence to:Shanye Gu , Email:shanye_gu@163.com Lai Jiang , Email:jianglai@xinhuamed.com.cn Cheng Huang , Email:chuang@shutcm.edu.cn Jiulin Du , Email:forestdu@ion.ac.cn;
J Mol Cell Biol, Volume 13, Issue 1, January 2021, 75-77,  https://doi.org/10.1093/jmcb/mjaa051

Targeted double-strand breaks (DSBs) in genomes can be introduced efficiently by endonucleases (Urnov et al., 2010; Jinek et al., 2012; Joung and Sander, 2013), including zinc-finger nucleases, transcription activator-like effector nucleases, and clustered regularly interspaced palindromic repeats (CRISPR)/Cas9. After DSBs, DNA repair is mainly via homology-directed repair (HDR) and/or non-homologous end joining (NHEJ) (Hustedt and Durocher, 2016). It was reported that genomic DNA replacement can be achieved via HDR at the site of DSBs in multiple organisms (Dickinson et al., 2013; Yang et al., 2013; Zu et al., 2013), but the efficiency is still not enough for general application, in particular for replacing long DNA fragment that is more than hundreds of base pairs (bps). As NHEJ is 10-fold more active than HDR at DSB sites (Mao et al., 2008), we speculated that NHEJ can be utilized to implement long genomic DNA replacement with high efficiency.


Volume 13, Issue 1
January 2021