Gene Targeting by Homology-Directed Repair in Rice Using a Geminivirus-Based CRISPR/Cas9 System
Dear Editor,Rice (Oryza sativa) is the staple food for more than half of the world”s population.Technologies enabling precise and efficient DNA knock-in or replacement,hereinafter referred to as KI,have the potential to revolutionize the generation of crops by precision molecular breeding.Clustered regularly interspaced short palindromic repeats (CRISPR)-associated Cas9 (CRISPR/Cas9)has recently emerged as a promising genome editing tool allowing precise genomic manipulation in rice and other crops.However,due to the prevalence of non-homologous end joining (NHEJ) over homology-directed repair (HDR) in the repair of CRISPR/Cas9-induced double-strand breaks (DSBs),this genome editing tool has been mostly used to generate random insertions and deletions (Indels) in precise genomic locations in plants (Cong et al.,2013;Feng et al.,2013;Miao et al.,2013;Shan et al.,2013;Ma et al.,2015;Xie et al.,2015;Gao et al.,2016).HDR-mediated DNA KI remains extremely challenging,partially due to the difficulty in delivering sufficient repair template,i.e.,donor DNA.Here,by combining CRISPR/Cas9 to produce DSBs and geminiviral vectors to deliver abundant donor DNA into rice cells,we have achieved up to 19.4％ targeted KI frequency in transgenic rice plants.
molecular breeding、transgenic rice、the world
10
S51;S5
This study was supported by the Chinese Academy of Sciences
2017-10-20（万方平台首次上网日期，不代表论文的发表时间）
共4页
1007-1010
