Efficient replacement of long DNA fragments via non-homologous end joining at non-coding regions

• 作者: Shanye Gu Jia Li Siyuan Li Jianbin Cao Jiwen Bu Yonggang Ren Wenjie Du Zhecong Chen Chufan Xu Mingcang Wang Lai Jiang Cheng Huang Jiulin Du
• 来源期刊: 分子细胞生物学报（英文版）
• 年，卷（期）: 2021,（1）
• 所属期刊栏目: Application Note
• 页码范围: 75-77
• 页数: 3页
• 语种: 英文
• DOI: 10.1093/jmcb/mjaa051

摘要

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 inter-spaced 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 partic-ular 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.