High-throughput continuous evolution of compact Cas9 variants targeting single-nucleotide-pyrimidine PAMs
PAM-containing sites in HEK293T cells. Mean±SEM is shown and reflects the average activity and standard error of= 3 independent biological replicates at the maximally edited position within each genomic site. The six sites that exhibited <1% base editing activity for either variant that were excluded in subsequent analyses are italicized.
Adenine base editing activity of eNme2-T.1- ABE8e or eNme2-T.2-ABE8e as a function of protospacer length at three different genomic sites in HEK293T cells. Each point represents the average of= 3 independent biological replicates observed for a given protospacer length at one genomic site, normalized to the protospacer length with the highest base editing activity for that site.
Nuclease activity of eNme2-C nuclease and eNme2-C.NR nuclease compared to SpRY nuclease and SpRY-HF1 nuclease at eight NCN/N4CN PAM-matched sites in HEK293T cells . For , Mean±SEM is shown and reflects the average activity and standard error of= 3 independent biological replicates measured at the maximally edited position within each given genomic site.
Extended Data Fig. 8 GUIDE-Seq identified off-targets of Nme2 variants compared to SpRY and SpRY-HF1.
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