Extended Data Fig. 7: Characterisation of CRISPRa-induced hPGCLCs.
From: Sequential enhancer state remodelling defines human germline competence and specification
a and b, FACS analysis of day 4 EBs generated from PreME of hESC lines harbouring the Dox-inducible CRISPRa transgene with the indicated sgRNA combinations. c, Immunofluorescence showing the co-expression of hPGCLC markers NANOS3-tdTomato, POU5F1 and SOX17 in hPGCLCs (yellow arrowheads) induced by CRISPRa in the absence of BMP4. White arrowheads indicate SOX17 single-positive cells (presumably DE). Representative results of 3 biological replicates. d, Induction of hPGCLCs from hESCs, PreME and ME with or without CRISPR-mediated activation of SOX17 and PRDM1 enhancers and promoters. FACS analysis of day 4 EBs shows that the co-activation of SOX17 and PRDM1 act synergistically with BMP4 to increase the efficiency of hPGCLC induction from hESCs and PreME, but not from ME. The appearance of the EBs under brightfield and tdTomato fluorescence filter are shown next to the corresponding FACS plots. Representative results of 3 independent experiments. e, Alluvial plots showing enhancer state transitions of hPGCLC-active enhancers in hESCs, PreME and ME.
