Genome editing mediated by SpCas9 variants with broad non-canonical PAM compatibility in plants

Streptococcus pyogenes Cas9 (SpCas9) is the most widely used genome editing tool in plants. The induced by SpCas9 strictly requires a canonical NGG protospacer-adjacent motif (PAM), significantly limiting its scope of application. Recently, five variants, SpCas9-NRRH, SpCas9-NRCH, SpCas9-NRTH, SpG, and SpRY, were developed to recognize non-canonical PAMs human cells. In this study, these variants engineered for plant editing, their targeted mutagenesis capabilities comprehensively examined at various PAM sites rice (Oryza sativa) stable transformation. Moreover, both cytosine base editors using rat APOBEC1 or APOBEC3a adenine directly evolved highly compatible TadA?-8e deaminase from variants. Our results demonstrated that variants-based readily generated conversions between C?G T?A target with transgenic lines. Collectively, toolbox study substantially expands SpCas9-mediated will greatly facilitate gene disruption precise