2026-04-17T05:00:23Zhttps://recercat.cat/oai/request

oai:recercat.cat:10230/481702025-12-12T02:24:21Zcom_2072_6col_2072_452952

00925njm 22002777a 4500 dc Bevacqua, Romina J. author Dai, Xiaoqing author Lam, Jonathan Y. author Gu, Xueying author Friedlander, Mollie S. H. author Tellez, Krissie author Miguel Escalada, Irene author Bonàs-Guarch, Silvia author Atla, Goutham author Zhao, Weichen author Kim, Seung Hyun author Dominguez, Antonia A. author Qi, Lei S. author Ferrer, Jorge author MacDonald, Patrick E. author Kim, Seung K. author 2021-07-14T07:01:05Z 2021-07-14T07:01:05Z 2021 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.) Diabetis Genètica Genòmica Proteïnes CRISPR-based genome editing in primary human pancreatic islet cells