【摘 要】
：
Dear Editor,rnThe clustered regularly interspaced short palindromic re-peats/CRISPR-associated nuclease 9(CRISPR/Cas9)sys-tem,since it was excavated,has been rapidly developed and sparked a revolution in the genome editing field.In principle,CRISPR/Cas9 s
【机 构】
：
State Key Laboratory of Rice Biology,China National Rice Research Institute,Chinese Academy of Agric
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Dear Editor,rnThe clustered regularly interspaced short palindromic re-peats/CRISPR-associated nuclease 9(CRISPR/Cas9)sys-tem,since it was excavated,has been rapidly developed and sparked a revolution in the genome editing field.In principle,CRISPR/Cas9 system relies on the recognition of specific loci on the genome,which is titled the protospacer adjacent motif(PAM).However,the canonical Streptococcus pyo-genes Cas9(SpCas9)nuclease only recognizes NGG or NAG PAMs,rendering an inherent obstacle in amplifying the application of CRISPR/Cas9 technology.In previous work,two main strategies were pursued to eliminate this constraint(Chen et al.,2019):one powerful pathway is mining SpCas9 protein orthologs,such as SaCas9 for NNGRRT PAMs and ScCas9 for NNG PAMs(Xu et al.,2020;Wang et al.,2020);another key approach is to generate SpCas9 variants,such as SpCas9-VQR and VRER for NGA and NGCG PAMs,SpCas9-NG for NG PAMs and xCas9 for NG,GAA and GAT PAMs.So far,although the potential scope of SpCas9 has been expanded in plant research,it is still insufficient for genome-wide arbitrary site targeting and editing.
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