Fig. 3.
Disruption of CCR5 in Jurkat T cells protects cells from HIV-1 infection via LentiCRISPR/SaCas9. a Mutation of CCR5 in Jurkat T cells was detected by T7E1 assay. Jurkat T cells were transduced with SaCas9/sgRNA-#6, #8 or control lentivirus with MOI of 40. After 3 days selection by puromycin, the cells were harvested for T7E1 assay. b Western blotting assessment of CCR5 expression on SaCas9/sgRNA-lentivirus edited Jurkat T cells. CCR5-modified or control Jurkat T cells in a were collected for western blotting assay with anti-CCR5 and anti-GAPDH antibodies. c DNA sequencing of CCR5 gene target loci in the modified Jurkat T cells. d The CCR5-sgRNA transduced Jurkat T cells attenuated HIV-1YU2 infection. The SaCas9/sgRNA-#6, #8 or control-treated Jurkat T cells were infected with R5-tropic HIV-1YU2 (MOI = 0.1). After 1, 3, 5, 7 days infection, cell culture medium was collected for p24 ELISA assay. Data were analyzed by unpaired t-test and error bars showed the mean ± SEM of three independent experiments (*p < 0.05; **p < 0.01; ***p < 0.001)