Fig. 2.

Single-stranded oligonucleotides mediated HDR editing attenuates large deletions. a Schematic for editing human T cells, HSPCs, and iPSCs with Cas9-gRNA RNPs and ssODN HDR donor templates. b Frequencies of small indels and HDR events after transfection of RNPs with or without ssODN donors in T cells, HSPCs, and iPSCs. Frequencies of indels (NHEJ) and HDR were determined by NGS and CRISPResso2 analysis 3 days after transfection. The numbers in the bars indicate the mean NHEJ or HDR efficiencies. c Frequencies of deletion indexes after RNP-ssODN editing in T cells, HSPCs, and iPSCs. The deletion indexes were determined by long-range PCR, ONT sequencing, alignment with references using Minimap2, and Samtools analysis. Error bars represent the mean ± SEM of n = 3–8 independent experiments. d Analysis of D100 in c. e Correlation of deletion indexes (DI) and deletion > 100 bp (D100) from data in c. The data in b, c, and d were statistically analyzed by a two-way ANOVA test. Adjusted p values were indicated. “ns” means no significance (p > 0.05)