Fig. 3: HDR-boosting modules achieve efficient precise gene editing at endogenous genomic loci in multiple human cell types.
From: Enhancing homology-directed repair efficiency with HDR-boosting modular ssDNA donor
a–d HDR efficiencies of HDR-boosting modular ssDNA donors and canonical ssDNA donors at the specified gene loci for EMX1, DNMT1, CXCR4, RUNX1, RNF2, and FANCF in HEK 293T cells (a), HeLa cells (b), U2OS cells (c), and K562 cells (d). e HDR efficiencies yielded with different concentrations of HDR-boosting modular ssDNA donors and canonical ssDNA donors at the FANCF site in HeLa cells. The digits denote the amount of ssDNA donors corresponding to 1 x 106 cells. f HDR efficiencies of ssDNA donors interrogating two endogenous gene sites simultaneously in HeLa cells. g HDR efficiencies of HDR-boosting modular ssDNA donors and canonical ssDNA donors at the FANCF site in hPB CD34+ cells. For all HDR efficiency-assessing experiments unless otherwise specified, 18 pmol Cas9 nuclease, 22 pmol gRNA and 6 pmol ssDNA donors corresponded to 2 × 105 cells. HDR efficiency was measured by NGS three days after electroporation. HDR efficiencies reflect the sequencing reads that contain the intended edit and do not contain indels among all treated cells (a–g). Values reflect n = 2 (a–g) independent electroporation replicates. The sequences of all gRNAs and ssDNA donors used are shown in Supplementary Data 1 and 2. Source data are provided as a Source Data file.
