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CRISPR-based genome editing in primary human pancreatic islet cells.

Nature communications | 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.

Pubmed ID: 33893274 RIS Download

Associated grants

  • Agency: NIDDK NIH HHS, United States
    Id: R01 DK107507
  • Agency: NIDDK NIH HHS, United States
    Id: R01 DK108817
  • Agency: NIDDK NIH HHS, United States
    Id: UC4 DK098085
  • Agency: NIDDK NIH HHS, United States
    Id: U01 DK123743
  • Agency: Medical Research Council, United Kingdom
    Id: MR/L02036X/1
  • Agency: NIDDK NIH HHS, United States
    Id: P30 DK116074
  • Agency: CIHR, Canada
    Id: 148451
  • Agency: NIDDK NIH HHS, United States
    Id: R01 DK126482
  • Agency: NIDDK NIH HHS, United States
    Id: R01 DK128932
  • Agency: Wellcome Trust, United Kingdom
    Id: WT101033

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