Fig. 3.

Upregulation of FOXP3 mRNA in Jurkat cells via targeting of selected promoter and enhancer regions. Jurkat cells were electroporated with plasmids encoding indicated single sgRNAs or a combination of sgRNAs in combination with the dCas9-VPR transcriptional activator and fold activation of FOXP3 gene expression was measured in reference to a control sample, electroporated with plasmids encoding random sgRNA (rnd) and dCas9-VPR. Each experiment was repeated 3 times. Cells were lysed and RNA isolated 48 h post electroporation. a Jurkat cells were electroporated with a combination of four sgRNAs targeting each region [Core (Fox 1–4), CNS1 (Fox 5–8), CNS2 (Fox 9–12), CNS3 (Fox 13–16), Cage1 (Fox 17–20) and Cage2 (Fox 21–24)] and FOXP3 gene expression was measured. b Jurkat cells were electroporated with each sgRNA (Fox 1–24) separately and the effect of each single sgRNA on FOXP3 gene expression was measured. c Jurkat cells were electroporated with sgRNAs 1, 14, 15, 17 and 18 separately and in combination and FOXP3 gene expression was measured