Fig. 4

Simple strategy to achieve spatiotemporal control of one or more gene knockouts based on the DIC system. a Overview of the spatiotemporal control of one or more gene knockouts based on the DIC system. On the basis of PFFs carrying TRE3G-SpCas9-T2A-tdTomato cassette at the Hipp11 locus, spatiotemporal gene knockout could be fulfilled only after transgene integration of spatial-specific-expressing rtTA and targeting interested-gene sgRNA expression cassette. b Outlined schematic for the establishment of cell line with spatiotemporal control of one or more gene knockouts based on the DIC system. c Schematic of the gene targeting strategy of GATA4, GATA6, and simultaneous GATA4 and GATA6 spatiotemporal gene knockouts. d Summary of cell screening experiments for GATA4, GATA6, and simultaneous GATA4 and GATA6 spatiotemporal gene knockouts