Fig. 4.

Knockout of TRAC and CD52 in human primary T cells. Four days after transfection of T cells with empty (No gRNA) or TRAC or CD52 gRNA-expressing pSpCas9-2A-GFP construct, the cells were analysed by a) IDAA, revealing a 1-bp insertion elicited by both gRNAs and no indels in empty pSpCas9-2A-GFP/No gRNA transfected cells or by b) flow cytometry for CD3 (TCR) and CD52 surface expression. This analysis showed an ~ 8% increase of cells in the gates representing background CD3 or CD52 expression levels, indicating that TCR and CD52 knockout was achieved in ~ 8% of the cells