Fig. 4: CRISPR/Cas9 targeting of a non-coding variant in the ABCC8-KCNJ11 locus impairs ABCC8 expression and function in primary human islets.
From: CRISPR-based genome editing in primary human pancreatic islet cells
A Schematic of the lenti-construct used for simultaneous expression of two sgRNAs, Cas9 and GFP (12,021 bp) in primary human islets. B Genome Browser tracks of the genomic context in the KCNJ11-ABCC8 locus, arcs representing high-confidence pcHi-C interactions in human islets, highlighting variant rs1002226 (chr11:17405617) associated with diabetes risk (black arrowhead): This variant maps to a CTCF site (blue) on a class I active enhancer (yellow line, and zoomed inset); sg_EK1 and sg_EK2 flanking rs1002226 (green arrows); Chromatin classes: active promoter (green); active enhancer (red); inactive enhancer (gray); inactive open chromatin (black); strong CTCF (blue). Accessible chromatin regions in human islets are shown by ATAC-seq, H3K27ac, and Mediator ChiP-seq. C ABCC8 mRNA is regulated by the rs1002226-containing enhancer in GFP+ pseudoislet cells, CRISPR-EK (green), CRISPR-Control (Black) (P = 0.0232; n = 4), while expression of KCNJ11, control genes (USH1C, NCR3LG1) in the same transcription activation domain, TAD (n = 3) and insulin (n = 4) were not modified. Data are presented as mean values +SD. D Measurement of KATP currents in GFP+ islet cells from CRISPR-EK (n = 26 cells, 3 replicates) and CRISPR-Control (n = 33, 3 replicates) (P < 0.0001). Data are presented as mean values ± SEM. Two-tailed t tests were used to generate P-values. *P < 0.05, **** P < 0.0001. Source data are provided as a Source Data file.
