Multigene Editing Via CRISPR/Cas9 Guided By a Single-sgRNA Seed in Arabidopsis

We report that a single-sgRNA seed is capable of guiding CRISPR/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 single sgRNA seed-induced 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.