CRISPR–Cas9 nucleases have transformed genome engineering, yet their application is often constrained by protospacer-adjacent motif (PAM) requirements. Staphylococcus aureus Cas9 (SaCas9) is particularly attractive for in vivo applications due to its compact size; however, its NNGRRT PAM limits targetable genomic sites. Here, we report KRH, a SaCas9 variant designed entirely from the wild-type enzyme through a fully computational point-mutation design workflow, UniDesign, without additional expe
Fully computational design of PAM-relaxed Staphylococcus aureus Cas9 with expanded targeting capability
Youcai Xiong·Xianan Huang·Li-Kuang Tsai·Xiaofeng Xia·Y Eugene Chen·Jie Xu·Jun Zhou·Shuang Chen·J. Zhang
