Supplementary Figure 7: Guide Swap is amenable to genome-scale screening in CD34+ HSPCs.
From: Guide Swap enables genome-scale pooled CRISPR–Cas9 screening in human primary cells
(a) CD33 (left) and CD45 (right) knockout efficiency in CD34 + HSPC from indicated donors. Gated on RFP + cells; two technical replicates in one experiment. mPB = mobilized peripheral blood. CB = cord blood. (b) Representative FACS plots of CD45 knockout in cord blood CD34 + CD90 + cells (top). Gating strategy for CD34 + CD90- and CD34 + CD90 + populations (bottom left). CD45 knockout efficiency in CB CD34 + CD90- and CD34 + CD90 + cells (bottom right). n = 2 technical replicates; representative of multiple gRNAs tested in two independent experiments with similar results. (c) CD34 + HSPC recovery 20 h post-electroporation, normalized to starting cell number. n = 8 technical replicates, data is presented as the mean. (d) Representative FACS plots of CD45RA and CD34 expression 7 and 14 days post-electroporation; two technical replicates in one experiment.
