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Fig. 2 | Retrovirology

Fig. 2

From: CCR5 editing by Staphylococcus aureus Cas9 in human primary CD4+ T cells and hematopoietic stem/progenitor cells promotes HIV-1 resistance and CD4+ T cell enrichment in humanized mice

Fig. 2

LentiCRISPR/SaCas9 mediated cleavage of CCR5 in TZM-bl cells against HIV-1 infection. a LentiCRISPR/SaCas9 editing of the CCR5 gene in TZM-bl cells. sgRNA-#6, #8 or control were transfected into TZM-bl cells delivered by lentiviral vector. Three days post transduction, genomic DNA was extracted and T7E1 assay was conducted. b Detection of CCR5 expression in TZM-bl cells by flow cytometry. c Sanger sequencing of CCR5 target loci in the TZM-bl cells modified by LentiCRISPR-SaCas9/sgRNA. d Disruption of CCR5 in TZM-bl cells could render cell resistance to HIV-1 infection. The cells were transduced with LentiCRISPR-SaCas9/sgRNA-#6, #8 or control. Then the modified cells were infected with HIV-1YU2 (MOI = 0.5). Luciferase reporter assay was performed to analyze infection efficiency. 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)

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Retrovirology

ISSN: 1742-4690

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