Fig. 4

CRISPR–Cas9 screen confirms CRC-TFs’ role in keratinocyte stem cell fate. a Schematic depicting pooled screening approach in Cas9-expressing neonatal HPEKs (nHPEK-Cas9) infected with sgRNA library (at day 0) followed by puromycin selection and collection of cell pellets at days 12, 19, 28, 33, 38 and 43 post infection after split. Abundance of sgRNA sequences was quantified by deep sequencing of the corresponding barcodes in the genomic DNA of the different cell pellets. b Gene level results of pooled screen at days 12, 19, 38, and 43, plotting z-score (Q1 or Q3) versus redundant siRNA activity (RSA Down or RSA Up). Q1 versus RSA Down at Days 12 and 19 identifies gene editing that reduces proliferation while Q3 versus RSA Up at Days 38 and 43 identifies gene editing that increases proliferation. The top 7 Core Regulatory Circuitry (CRC) TFs (from Fig. 3d) are highlighted and marked as associated with either HSCP (blue), LSCP (red), or “neutral” (green)