Multiplexed CRISPR gene editing in primary human islet cells with Cas9 ribonucleoprotein.

Techniques in genetics biology experimental methods cell biology human genetics

Journal

iScience
ISSN: 2589-0042
Titre abrégé: iScience
Pays: United States
ID NLM: 101724038

Informations de publication

Date de publication:
19 Jan 2024
Historique:
received: 26 06 2023
revised: 27 10 2023
accepted: 05 12 2023
medline: 11 1 2024
pubmed: 11 1 2024
entrez: 11 1 2024
Statut: epublish

Résumé

Successful genome editing in primary human islets could reveal features of the genetic regulatory landscape underlying β cell function and diabetes risk. Here, we describe a CRISPR-based strategy to interrogate functions of predicted regulatory DNA elements using electroporation of a complex of Cas9 ribonucleoprotein (Cas9 RNP) and guide RNAs into primary human islet cells. We successfully targeted coding regions including the

Identifiants

DOI: 10.1016/j.isci.2023.108693 PMID: 38205242 PMC: PMC10777115
pubmed: 38205242
doi: 10.1016/j.isci.2023.108693
pii: S2589-0042(23)02770-0
pmc: PMC10777115
doi:

Types de publication

Journal Article

Langues

eng

Pagination

108693

Informations de copyright

© 2023 The Author(s).

Déclaration de conflit d'intérêts

A.M. is a co-founder of Arsenal Biosciences, Function Bio, Spotlight Therapeutics, and Survey Genomics, serves on the boards of directors at Function Bio, Spotlight Therapeutics, and Survey Genomics, is a member of the scientific advisory boards of Arsenal Biosciences, Function Bio, Spotlight Therapeutics, Survey Genomics, NewLimit, Amgen, Tenaya, and Lightcast, owns stock in Arsenal Biosciences, Function Bio, Spotlight Therapeutics, NewLimit, Survey Genomics, Tenaya, and Lightcast, and has received fees from Arsenal Biosciences, Spotlight Therapeutics, NewLimit, 23andMe, PACT Pharma, Juno Therapeutics, Tenaya, Lightcast, Trizell, Vertex, Merck, Amgen, Genentech, AlphaSights, Rupert Case Management, Bernstein, GLG, ClearView Healthcare Partners, and ALDA. A.M. is an investor in and informal advisor to Offline Ventures and a client of EPIQ. The Marson laboratory has received research support from Juno Therapeutics, Epinomics, Sanofi, GlaxoSmithKline, Gilead, and Anthem. A.L.G.’s spouse holds stock options in Roche and is an employee of Genentech.

Auteurs

Romina J Bevacqua (RJ)

Department of Developmental Biology, Stanford University School of Medicine, Stanford, CA 94305, USA.
Diabetes, Obesity and Metabolism Institute (DOMI), Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.

Weichen Zhao (W)

Department of Developmental Biology, Stanford University School of Medicine, Stanford, CA 94305, USA.

Emilio Merheb (E)

Diabetes, Obesity and Metabolism Institute (DOMI), Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.

Seung Hyun Kim (SH)

Department of Developmental Biology, Stanford University School of Medicine, Stanford, CA 94305, USA.

Alexander Marson (A)

Gladstone-UCSF Institute of Genomic Immunology and Northern California JDRF Center of Excellence, University of California at San Francisco, San Francisco, CA 94158, USA.
Department of Medicine, University of California, San Francisco, San Francisco, CA 94143, USA.
Diabetes Center, University of California, San Francisco, San Francisco, CA 94143, USA.
Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, CA 94143, USA.

Anna L Gloyn (AL)

Department of Pediatrics (Endocrinology) and of Genetics, Stanford University School of Medicine, Stanford, CA 94305, USA.
Stanford Diabetes Research Center, Stanford University School of Medicine, Stanford, CA 94305, USA.

Seung K Kim (SK)

Department of Developmental Biology, Stanford University School of Medicine, Stanford, CA 94305, USA.
Departments of Medicine and of Pediatrics, Stanford University School of Medicine, Stanford, CA 94305, USA.
Stanford Diabetes Research Center, Stanford University School of Medicine, Stanford, CA 94305, USA.
Northern California JDRF Center of Excellence, Stanford University School of Medicine, Stanford, CA 94305, USA.

Classifications MeSH