CRISPR-based genome editing in primary human pancreatic islet cells

  • Nat Commun. 2021 Apr 23;12(1):2397. doi: 10.1038/s41467-021-22651-w.
Romina J Bevacqua  1 ,  Xiaoqing Dai  2 ,  Jonathan Y Lam  1 ,  Xueying Gu  1 ,  Mollie S H Friedlander  1 ,  Krissie Tellez  1 ,  Irene Miguel-Escalada  3  4 ,  Silvia Bonàs-Guarch  3  4 ,  Goutham Atla  3  4 ,  Weichen Zhao  1 ,  Seung Hyun Kim  1 ,  Antonia A Dominguez  5  6 ,  Lei S Qi  5  6  7 ,  Jorge Ferrer  3  4  8 ,  Patrick E MacDonald  2 ,  Seung K Kim  9  10  11  12
Affiliations
  • 1. Department of Developmental Biology, Stanford University School of Medicine, Stanford, CA, USA.
  • 2. Alberta Diabetes Institute and Department of Pharmacology, University of Alberta, Edmonton, AB, Canada.
  • 3. Bioinformatics and Genomics Program, Centre for Genomic Regulation (CRG), The Barcelona Institute of Science and Technology (BIST), Barcelona, Spain.
  • 4. Centro de Investigación Biomédica en Red de Diabetes y Enfermedades Metabólicas Asociadas (CIBERDEM), Barcelona, Spain.
  • 5. Department of Bioengineering, Stanford University, Stanford, CA, USA.
  • 6. Department of Chemical and Systems Biology, Stanford University, Stanford, CA, USA.
  • 7. Chem-H, Stanford University, Stanford, CA, USA.
  • 8. Section of Genetics and Genomics, Imperial College London, London, UK.
  • 9. Department of Developmental Biology, Stanford University School of Medicine, Stanford, CA, USA. [email protected].
  • 10. Department of Medicine (Endocrinology), Stanford University School of Medicine, Stanford, CA, USA. [email protected].
  • 11. Northern California JDRF Center of Excellence, Stanford University School of Medicine, Stanford, CA, USA. [email protected].
  • 12. Stanford Diabetes Research Center, Stanford University School of Medicine, Stanford, CA, USA. [email protected].
PMID: 33893274 DOI: 10.1038/s41467-021-22651-w
Abstract

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.

Products