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

摘要

We report that a solo single-guide RNA(sg RNA) seed is capable of guiding Clustered Regularly Interspaced Short Palindromic Repeats(CRISPR)/CRISPRàassociated 9(CRISRP/Cas_9) to simultaneously edit multiple genes At RPL_(10)A, At RPL_(10)B and At RPL_(10)C in Arabidopsis. Our results also demonstrate that it is possible to use CRISPR/Cas_9 technology to create At RPL_(10) triple mutants which otherwise cannot be generated by conventional genetic crossing. Compared to other conventional multiplex CRISPR/Cas systems, a single sg RNA 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.