CRISPR/Cas9-mediated knockout of clinically relevant alloantigenes in human primary T cells

摘要

FACS isolation of gRNA/Cas9-expressing 293 T cells. Three days after transfection of 293 T cells with no construct or TRAC or CD52 gRNA-expressing pSpCas9-2A-GFP construct, the cells were subjected to flow cytometry. Analysis of non-transfected cells was used to define the autofluorescence level of the cells in the GFP channel (marked by a bar in the graphs). Next, cells with high GFP expression levels (population 4) were bulk FACS isolated