Exploring the deletion landscape of <i>S. aureus</i> Cas9 with SABER.

ABSTRACT: Profiling tolerated amino acid deletions in proteins can elucidate structure-function relationships, reconstruct intermediate stages in protein evolution, and be used to engineer minimized versions of proteins with size-sensitive biotechnology applications. Despite advances in deletion library construction techniques over the past several decades, there are presently few methods available that are simultaneously efficient, precise, and easy to implement. Here we present SABER, a novel approach which utilizes SpRYCas9, a near-PAMless engineered SpCas9 variant, as a molecular biological tool for building deletion libraries with unprecedented speed and ease. We applied this technique to the small and structurally divergent Cas9 from Staphylococcus aureus (SaCas9) and mapped the set of

SUBMITTER: Plebanek AJ 

PROVIDER: S-EPMC12363808 | biostudies-literature | 2025 Aug

REPOSITORIES: biostudies-literature