Supplementary Figure 3: Efficient CD45 knockout using lenti gRNA and RNPs of varied gRNA targets and formats.
From: Guide Swap enables genome-scale pooled CRISPR–Cas9 screening in human primary cells
(a)-(c) are representative of multiple lenti gRNAs tested in two independent experiments with similar results. (a) Flow cytometry analysis of CD45 knockout efficiency 4 days post-electroporation of transduced CD34 + HSPC with indicated RNPs. Lentiviral gRNA-expressing cells were gated using RFP. n = 2 technical replicates. The SRPK1 and ZNRF3 gRNAs were unmodified. All other gRNAs were chemically modified. (b) Flow cytometry analysis of CD45 knockout efficiency 4 days post-electroporation of transduced CD34 + HSPC with indicated RNPs (dg = synthetic split gRNA; sg = synthetic single gRNA). Lenti gRNA-expressing cells were gated using the RFP marker. n = 2 technical replicates. (c) Representative FACS plots of CD33 and CD45 expression 4 days post-electroporation with indicated RNPs. Lenti gRNA-expressing cells were gated using the RFP marker.
