<i>CCR5</i> gene disruption by RNA-guided Cas9 endonuclease in transduced CEM ss-CCR5 cells.

Abstract

<p>A. Cell surface expression of CCR5 on CEMss-CCR5 cells co-transduced with lentiviral vectors expressing Cas9-HA-NLS and one of sgRNA (CR2 or GF1) as compared to mock-transduced CEMss-CCR5 cells. B. FSC and SSC analysis mock-transduced CEMss-CCR5 cells and CEMss-CCR5 cells transduced with lentiviral vectors expressing Cas9-HA-NLS fusion protein and sgRNAs CR2 or GF1, respectively. Percentages of gated (live) cells are shown at the lower and right corner of the figures. C. <i>CCR5</i> gene disruption analysis in mock-transduced CEMss-CCR5 cells or CEMss-CCR5 cells co-transduced with lentiviral vectors expressing Cas9-HA-NLS and one of sgRNAs (CR2 or GF1) by T7EI assay. D. Mean and median values of fluorescence intensity of cell surface expression of CD4, CXCR4 and CCR5 in sorted CCR5-/CR2 and CCR5+/GF1 cells as compared to parental CEMss-CCR5 cells. E. Relative fluorescent intensity of DDAO-labeled CCR5-/CR2 and CCR5+/GF1 cells as compared to parental CEMss-CCR5 cells at 0, 12, 24 and 36 hours post culture.</p