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


Journal

bioRxiv : the preprint server for biology
Titre abrégé: bioRxiv
Pays: United States
ID NLM: 101680187

Informations de publication

Date de publication:
17 Sep 2023
Historique:
pubmed: 25 9 2023
medline: 25 9 2023
entrez: 25 9 2023
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

pubmed: 37745551
doi: 10.1101/2023.09.16.558090
pmc: PMC10516051
pii:
doi:

Types de publication

Preprint

Langues

eng

Subventions

Organisme : NIDDK NIH HHS
ID : R01 DK107507
Pays : United States
Organisme : NIDDK NIH HHS
ID : R01 DK108817
Pays : United States
Organisme : NIDDK NIH HHS
ID : UC4 DK098085
Pays : United States
Organisme : Wellcome Trust
Pays : United Kingdom
Organisme : NIDDK NIH HHS
ID : UM1 DK126185
Pays : United States
Organisme : NIDDK NIH HHS
ID : U01 DK123743
Pays : United States
Organisme : NIDDK NIH HHS
ID : P30 DK116074
Pays : United States

Déclaration de conflit d'intérêts

Declaration of interests R.J.B., S.H.K., W.Z., and S.K.K. declare no competing interests. A.M. is a co-founder of Arsenal Biosciences, Spotlight Therapeutics, and Survey Genomics, serves on the boards of directors at Spotlight Therapeutics and Survey Genomics, is a board observer (and former member of the board of directors) at Arsenal Biosciences, is a member of the scientific advisory boards of Arsenal Biosciences, Spotlight Therapeutics, Survey Genomics, NewLimit, Amgen, Tenaya, and Lightcast. A.M. owns stock in Arsenal Biosciences, Spotlight Therapeutics, NewLimit, Survey Genomics, PACT Pharma, Tenaya, and Lightcast and has received fees from Arsenal Biosciences, Spotlight Therapeutics, NewLimit, Survey Genomics, 23andMe, PACT Pharma, Juno Therapeutics, Tenaya, Lightcast, Trizell, Vertex, Merck, Amgen, Genentech, AlphaSights, Rupert Case Management, Bernstein, 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.
Mount Sinai Regenerative Biology and Stem Cell Institute, New York, NY, United States.

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, 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.
Departments of Medicine and of Pediatrics, 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