CRISPR-based genome editing in primary human pancreatic islet cells

• Published in: Nature communications 2021-04, Vol.12 (1), p.2397-2397, Article 2397
• Main Authors: Bevacqua, Romina J., Dai, Xiaoqing, Lam, Jonathan Y., Gu, Xueying, Friedlander, Mollie S. H., Tellez, Krissie, Miguel-Escalada, Irene, Bonàs-Guarch, Silvia, Atla, Goutham, Zhao, Weichen, Kim, Seung Hyun, Dominguez, Antonia A., Qi, Lei S., Ferrer, Jorge, MacDonald, Patrick E., Kim, Seung K.
• Format: Article
• Language: English
• Subjects: 13; 13/106; 38; 42/41; 42/44; 631/337/4041/3196; 692/699/2743/137; Base Sequence; Beta cells; CRISPR; CRISPR-Cas Systems; Deoxyribonucleic acid; Diabetes; Diabetes mellitus; Diabetes mellitus (non-insulin dependent); Diabetes Mellitus, Type 2 - genetics; DNA; Editing; Exons; Gene Editing - methods; Gene Expression Regulation; Gene targeting; Genome editing; Genomes; Health risks; Homeodomain Proteins - genetics; Humanities and Social Sciences; Humans; Insulin-Secreting Cells - cytology; Insulin-Secreting Cells - metabolism; Islet cells; Islets of Langerhans - cytology; Islets of Langerhans - metabolism; Models, Genetic; multidisciplinary; Pancreas; Pathogenesis; Potassium channels (inwardly-rectifying); Potassium Channels, Inwardly Rectifying - genetics; Proteins; Regulators; Regulatory sequences; Science; Science (multidisciplinary); SIX gene family; Trans-Activators - genetics
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-021-22651-w

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 K ATP 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. The editing of primary human islets could provide insight into diabetes pathogenesis. Here the authors use CRISPR-Cas9 to target regulatory elements associated with T2D susceptibility.