Fig. 1: In vitro CRISPR/Cas9-based screen to identify determinants for HSPC expansion. | Leukemia

Fig. 1: In vitro CRISPR/Cas9-based screen to identify determinants for HSPC expansion.

From: In vivo CRISPR/Cas9-mediated screen reveals a critical function of TFDP1 and E2F4 transcription factors in hematopoiesis

Fig. 1: In vitro CRISPR/Cas9-based screen to identify determinants for HSPC expansion.

A Scheme of CRISPR/Cas9-based screen in mouse HSPCs. Cas9-expressing HSPCs were isolated from R26-Cas9iGFP mice, mixed with C57BL/6 (WT) HSPCs at a ratio of 1:1, and activated for one day. The activated HSPCs were infected with lentiviral particles expressing specific sgRNA in 96-well plates. These cells were cultured and analyzed on day two and seven after infection by flow cytometry. The survival/proliferation (sur/pro) score of the pre-gated HSPCs was defined as indicated. B Representative FACS analysis of the frequency of GFP+ (Cas9+) cells within mCherry+ (sgRNA+) or mCherry- (sgRNA-) HSPCs two (top) and seven days (bottom) post transduction. In each column, the indicated sgRNA was used. C Correlation of survival/proliferation scores of two independent experiments at the indicated time points (n = 2 biological replicates). Genes with a survival/proliferation score <−1 are highlighted.

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