Fig. 2

High HDR efficiency is achieved in 293 T cells by double cut HDR donor even with short HA. a Schematic outline of pD-mCherry-sg (double cut HDR donor) with HA in the range of 0–1500 bp in length. The red triangle indicates a sgRNA target sequence. The left arm is marked as yellow and the right arm as blue. b Determination of the HDR efficiency by FACS. 293 T-reporter cells were analyzed one week after co-transfection of Cas9, sgRNA1, together with either pD-mCherry or pD-mCherry-sg. The percentages of mCherry⁺ cells represent the HDR efficiencies. c Effects of HA length of conventional and double cut donors on HDR efficiency. n = 3 biological replicates; error bars represent S.E.M. Significance was calculated using the Student’s paired t-test: *P ≤ 0.05; **P ≤ 0.01; ***P ≤ 0.001; ns not significant