Extended Data Fig. 8: GUIDE-Seq identified off-targets of Nme2 variants compared to SpRY and SpRY-HF1. | Nature Biotechnology

Extended Data Fig. 8: GUIDE-Seq identified off-targets of Nme2 variants compared to SpRY and SpRY-HF1.

From: High-throughput continuous evolution of compact Cas9 variants targeting single-nucleotide-pyrimidine PAMs

Extended Data Fig. 8

(a) On-target indel formation of wild-type Nme2 nuclease, eNme2-C nuclease, and eNme2-C.NR nuclease compared to SpRY nuclease and SpRY-HF1 nuclease at each of the four protospacer-matched sites that were subsequently evaluated in GUIDE-Seq. Each bar represents the observed indel formation of one replicate in U2OS cells. (b-e) GUIDE-Seq identified off-targets and associated read counts for Nme2 variants (top) or SpRY variants (bottom) at Site 3 (b), Site 4 (c), Site 5 (d), and Site 6 (e). The on-target protospacer is marked by a black dot for each site. Off-target thresholds were set at 8 mismatches with and NNN PAM for SpRY variants or 11 mismatches with an NNNNNN PAM for Nme2 variants).

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