CRISPR-based genome editing in primary human pancreatic islet cells

Gene targeting studies in primary human islets could advance our understanding of mechanisms driving diabetes pathogenesis. Here, we demonstrate successful genome editing in primary human islets using clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein 9 (Cas9). CRISPR-based targeting efficiently mutated protein-coding exons, resulting in acute loss of islet β-cell regulators, like the transcription factor PDX1 and the KATP channel subunit KIR6.2, accompanied by impaired β-cell regulation and function. CRISPR targeting of non-coding DNA harboring type 2 diabetes (T2D) risk variants revealed changes in ABCC8, SIX2 and SIX3 expression, and impaired β-cell function, thereby linking regulatory elements in these target genes to T2D genetic susceptibility. Advances here establish a paradigm for genetic studies in human islet cells, and reveal regulatory and genetic mechanisms linking non-coding variants to human diabetes risk.We gratefully acknowledge organ donors and their families, Canadian organ procurement organizations, particularly the Human Organ Procurement and Exchange (HOPE) program and the Trillium Gift of Life Network, and islet procurement through the Alberta Diabetes Institute Islet Core, Integrated Islet Distribution Program (U.S. NIH UC4 DK098085). R.J.B. was supported by a postdoctoral fellowship from JDRF (3-PDF-2018-584-A-N) and is on leave from the Animal Biotechnology Laboratory, Facultad de Agronomía, Universidad de Buenos Aires/INPA CONICET, CABA, Argentina. Work in the CRG and ICL was funded by the Wellcome Trust (WT101033), Medical Research Council (MR/L02036X/1), European Research Council Advanced Grant (789055), Ministerio de Ciencia e Innovación (RTI2018-095666-B-I00) and Marató TV3 #201611. Work in the University of Alberta was supported by a Foundation Grant from the Canadian Institutes of Health Research (CIHR: 148451, MacDonald). Work in the Kim lab was supported by NIH awards (R01 DK107507; R01 DK108817; U01 DK123743 to S.K.K.), and JDRF Northern California Center of Excellence (to S.K.K. and M. Hebrok). Work here was also supported by NIH grant P30 DK116074 (S.K.K.