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. 2022 Dec 2;119(49):e2204259119. doi: 10.1073/pnas.2204259119

Fig. 3.

Fig. 3.

PRDM9-Cas9 fusion protein displays increased HDR efficiency and HDR:indel ratio across multiple genomic sites. (A) HDR frequency and HDR:indel ratio measured by NGS at site C7 (intergenic) in HEK293T cells transfected with plasmids expressing fusion protein and sgRNA along with an ssODN template with 50-nt homology arms. (B) HDR frequency and HDR:indel ratio measured by NGS at site C9 (exon 5 of LDLR) in HEK293T cells transfected with fusion protein and sgRNA along with an ssODN template with 50-nt homology arms. (C) HDR frequency measured by NGS at 10 genomic loci in HEK293T cells transfected with PRDM9-Cas9 and sgRNA with an ssODN template. (D) HDR:indel ratio measured by NGS at 10 genomic loci in HEK293T cells transfected with PRDM9-Cas9 and sgRNA with an ssODN template. (E) Off-target activity of PRDM9-Cas9 measured by NGS at seven potential off-target sites predicted from two sgRNAs. For A–E, data represent mean ± SD (n = 3). *P < 0.05, **P < 0.01, and ***P < 0.001, determined by Student’s two-tailed t test.