Fig. 5

Evaluation of multiple symmetric and asymmetric ssODN donor designs used in combination with different CRISPR-Cas systems. a Various types of ssODN donor templates tested. Each single-strand DNA donor is complementary to either the target strand (and hence is of the non-target strand sequence and is denoted “NT”) or the non-target strand (and hence is of the target strand sequence and is denoted “T”). The NT ssODN donor is colored red, while the T ssODN donor is colored blue. The green box between the homology arms indicates the restriction site to be integrated into the target genomic locus. The 37/77 T ssODN has previously been found to be optimal for SpCas9-induced HDR [31]. b, c Extent of precise gene editing by SpCas9, AsCpf1, and LbCpf1 at the b CACNA1D or c PPP1R12C locus. Cells were harvested 72 h after transfection and the gene targeting efficiencies were determined by Illumina deep sequencing. Data represent mean ± standard error of the mean (s.e.m.; n ≥ 4). *P < 0.05, **P < 0.01, ***P < 0.001, N.S. not significant; Student’s t-test