GSE125959
A positive, growth-based PAM screen reveals non-canonical motifs recognized by the S. pyogenes Cas9
We report non-canonical PAM sequences for SpyCas9 that were revealed from a novel growth-based PAM screen in E. coli. Overall design: Next-generation sequencing was used to identify enriched PAM sequences from a 5N PAM library in E. coli. The library with and without IPTG induction was sent for sequencing to determine enrichment. NGS was also used to evaluate indel formation in U2OS cells after targeting by SpyCas9.