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. 2021 Apr 23;12:2397. doi: 10.1038/s41467-021-22651-w

Fig. 6. CRISPR/dCas9 activation of a putative enhancer element in the SIX2-SIX3 locus in primary human islets.

Fig. 6

A Genome browser tracks highlighting the putative SIX2-SIX3 enhancer element (SIXE) with variants previously linked to increased risk of fasting glucose hyperglycemia and T2D (T2D-SNPs, black arrowheads): These variants map to an active enhancer within an enhancer cluster (yellow line); sgRNAs used for CRISPRa: SIXE-A1, SIXE-A2. Chromatin classes: active enhancer (red); Accessible chromatin regions in human islets are shown by ATAC-seq, H3K27ac, and Mediator ChiP-seq. B Schematics of the lenti-construct used for simultaneous expression of two sgRNAs, VPR, dCas9 and GFP (13,728 bp) in primary human islets. C–D mRNA levels of (C) SIX2 (P = 0.042; n = 6 independent donors), SIX3 (P = 0.0328; n = 6 independent donors), PRKCE (n = 3 independent donors) and (D) INS (n = 5 independent donors), in GFP+ cells after CRISPRa of SIXE (CRISPRa-SIXE). Data are presented as mean values ± SD. Two-tailed t tests were used to generate P-values. *P < 0.05. Source data are provided as a Source Data file.