Fig. 3

The optimized sgRNA structure is superior to the original version. a CCR5 knockout efficiency was determined for the indicated sgRNAs targeting CCR5 with either an optimized sgRNA structure or the original structure. The knockout efficiency was determined in the same way as in Fig. 1b. The raw data are sown in Figure S6 in Additional file 1. b CD4 knockout efficiency was determined for the indicated sgRNAs targeting the CD4 gene, with two versions of the sgRNA structure in Jurkat cells. Cells were analyzed for CD4 expression by flow cytometry 72 hours after transfection. The raw data are shown in Figure S7 in Additional file 1. c T→C and T→G mutations are superior to the T→A mutation. Eleven sgRNAs targeting CCR5 were randomly selected. The knockout efficiency of sgRNAs with different mutations at position 4 in the sequence of continuous Ts were determined as in Fig. 1c. The raw data are shown in Figure S9 in Additional file 1. The graphs represent biological repeats from one of three independent experiments with similar results, shown as mean ± standard deviation (n = 3). Significance was calculated using Student's t-test: *P < 0.05; **P < 0.01; ***P < 0.005; ****P < 0.001