摘要
We report that a solo single-guide RNA （sgRNA） seed is capable of guiding Clustered Regularly Interspaced Short Palindromic Repeats （CRISPR）/CRISPR -associated 9 （CRISRP/Casg） to simultaneously edit multiple genes AtRPL10A, AtRPL10B and AtRPL10C in Arabidopsis. Our results also demonstrate that it is possible to use CRISPR/Cas9 technology to create AtRPLlo triple mutants which otherwise cannot be generated by conventional genetic crossing. Compared to other conventional multiplex CRISPR/Cas systems, a single sgRNA seed has the advantage of reducing off-target gene-editing. Such a gene editing system might be also applicable to modify other homologous genes, or even less-homologous sequences for multiple gene-editing in plants and other organisms.
We report that a solo single-guide RNA （sgRNA） seed is capable of guiding Clustered Regularly Interspaced Short Palindromic Repeats （CRISPR）/CRISPR -associated 9 （CRISRP/Casg） to simultaneously edit multiple genes AtRPL10A, AtRPL10B and AtRPL10C in Arabidopsis. Our results also demonstrate that it is possible to use CRISPR/Cas9 technology to create AtRPLlo triple mutants which otherwise cannot be generated by conventional genetic crossing. Compared to other conventional multiplex CRISPR/Cas systems, a single sgRNA seed has the advantage of reducing off-target gene-editing. Such a gene editing system might be also applicable to modify other homologous genes, or even less-homologous sequences for multiple gene-editing in plants and other organisms.
基金
supported by grants from the National Science Foundation of China(NSFC,31200913)
the Ministry of Agriculture of the People’s Republic of China National Transgenic Program(2016ZX08009001-004)
the China Scholarship Council(201709645003)
NSFC(31370180,31601765)
the Zhejiang Provincial Natural Science Foundation(LY15C140006,LY14C010005)
