Supplementary Figure 5: Guide Swap is amenable to genome-scale screening.
From: Guide Swap enables genome-scale pooled CRISPR–Cas9 screening in human primary cells
(a) CD34 + HSPC were transduced with the indicated lenti gRNAs, then electroporated with indicated amounts of RNP where X µg RNP = X µg Cas9 + X µg nt_A gRNA (1:1 crRNA:tracrRNA). CD45 expression was assessed by flow cytometry. Lenti gRNA-expressing cells were gated using RFP. n = 2 technical replicates. (b) Same as above except in T cells. CD45 (left) and CXCR4 (right) expression were assessed by flow cytometry. (c) The indicated numbers of transduced HSPC were pelleted and resuspended in 20 µL for electroporation. CD45 knockout was assessed by flow cytometry 4 days post-electroporation. Representative of multiple gRNAs tested in two independent experiments with similar results.
