Figure 7. Higher efficiency of HR-mediated gene targeting in NHEJ-deficient cells using different length of homologous arms.

Vector donors containing short homologous arms were used for BmTUDOR-SN gene editing of EGFP. Donors (300 ng/well each), pie2FW-Cas9 (300 ng/well), and Tend-gRNA expression vector (300 ng/well) were transfected into the indicated cells (1 × 10⁵ cells/well). Genomic DNA PCR was performed at 7 days post transfection of the donors as indicated. The gel images were analyzed by Image J to quantify the PCR products, with normalization to the reference bands. The PCR product from N4-SID1-Cas9 cells transfected with indicated donors was set as 1 fold. The agarose gel images were representatives from repeated three independent experiments. The numbers below the PCR bands represent mean fold ± S. D. from the three repeats.