Multigene editing via CRISPR/Cas9 guided by a single‐sgRNA seed in Arabidopsis

Summary 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/Cas9) 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 AtRPL10 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. A single‐sgRNA seed is capable of guiding CRISPR/Cas9 to simultaneously edit multiple genes AtRPL10A, AtRPL10B and AtRPL10C in Arabidopsis. Our results imply that such a multi‐gene editing system might be also applicable to modifying other homologous genes or even less‐homologous sequences in plants and other organisms.