Effect of deletion of ZHX3 on gene expression in human pancreatic beta cells EndoC-BH1
ZHX3 is a transcriptional factor whose role in human pancreatic beta cells is not well understood. In attempt to identify the genes that ZHX3 regulates in beta cells, we KO ZHX3 using CRISPR in EndoC-ßH1 cells and performed RNA-Seq. We have only identified 36 and 31 genes that were significantly up and downregulated respectively. This is in concordance with our functional data whereby the KO of ZHX3 in EndoC-ßH1 did not affect its insulin secretory function. Overall, we conclude that the KO of ZHX3 did not result in major perturbations in EndoC-ßH1 cells. Overall design: ZHX3 was deleted using the CRISPR-Cas9 system with one gRNA targeting the first coding exon of ZHX3 (sgZHX3-3). EndoC-ßH1 cells were transduced with either empty lentiCRISPRv2 plasmids or lentiCRISPRv2 plasmids containing sgZHX3-3. The cells were then selected with antibiotics and cultured as stable lines.