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Fig. 6 | Genome Biology

Fig. 6

From: Identification of X-chromosomal genes that drive sex differences in embryonic stem cells through a hierarchical CRISPR screening approach

Fig. 6

Knock-down of Dusp9 and Klhl13 in female mESCs leads to a shift towards the male pluripotency phenotype. a Catalytically dead Cas9 (blue) and the KRAB repressor domain (red) are each fused to one component of the PYL1-ABI system (gray), which dimerizes upon treatment with abscisic acid (ABA), resulting in gene repression. b CRISPR multiguide plasmid used for expression of three different sgRNAs targeting a specific gene. Each sgRNA is expressed under a different Pol III promoter, as indicated. c–e 1.8 female wildtype mESCs stably expressing the CRISPRi system shown in a were transduced with vectors expressing sgRNAs targeting Dusp9 (yellow), Klhl13 (blue), or a non-targeting control construct (NTC; black). Expression of each target gene (c), five MAPK target genes (d), and five pluripotency factors (e) was quantified by qPCR in cells expressing the respective sgRNAs or NTCs, as indicated. Bars represent the mean of 3 biological replicates, gray dots the individual measurements. Cells were treated with abscisic acid (ABA) for 5 days prior to cell harvesting for phenotypic assessment. *p < 0.05 two-tailed paired Student’s t test comparing gene-specific sgRNAs and NTCs are indicated

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