EpiCRISPR targeted methylation of Arx gene initiates transient switch of mouse pancreatic alpha to insulin-producing cells.


Journal

Frontiers in endocrinology
ISSN: 1664-2392
Titre abrégé: Front Endocrinol (Lausanne)
Pays: Switzerland
ID NLM: 101555782

Informations de publication

Date de publication:
2023
Historique:
received: 30 12 2022
accepted: 24 02 2023
medline: 4 4 2023
entrez: 3 4 2023
pubmed: 4 4 2023
Statut: epublish

Résumé

Beta cell dysfunction by loss of beta cell identity, dedifferentiation, and the presence of polyhormonal cells are main characteristics of diabetes. The straightforward strategy for curing diabetes implies reestablishment of pancreatic beta cell function by beta cell replacement therapy. Aristaless-related homeobox (Arx) gene encodes protein which plays an important role in the development of pancreatic alpha cells and is a main target for changing alpha cell identity. In this study we used CRISPR/dCas9-based epigenetic tools for targeted hypermethylation of Arx gene promoter and its subsequent suppression in mouse pancreatic αTC1-6 cell line. Bisulfite sequencing and methylation profiling revealed that the dCas9-Dnmt3a3L-KRAB single chain fusion constructs (EpiCRISPR) was the most efficient. Epigenetic silencing of In conclusion, we successfully triggered a direct, transient switch of pancreatic alpha to insulin-producing cells opening a future research on promising therapeutic avenue for diabetes management.

Identifiants

pubmed: 37008919
doi: 10.3389/fendo.2023.1134478
pmc: PMC10063207
doi:

Substances chimiques

Transcription Factors 0
Homeodomain Proteins 0
Insulin 0
ARX protein, mouse 0

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

1134478

Informations de copyright

Copyright © 2023 Đorđević, Stepper, Feuerstein-Akgoz, Gerhauser, Paunović, Tolić, Rajić, Dinić, Uskoković, Grdović, Mihailović, Jurkowska, Jurkowski, Jovanović and Vidaković.

Déclaration de conflit d'intérêts

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

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Auteurs

Marija Đorđević (M)

Department of Molecular Biology, Institute for Biological Research "Siniša Stanković" - National Institute of Republic of Serbia, University of Belgrade, Belgrade, Serbia.

Peter Stepper (P)

Institute of Biochemistry and Technical Biochemistry, University of Stuttgart, Stuttgart, Germany.

Clarissa Feuerstein-Akgoz (C)

Division of Epigenomics and Cancer Risk Factors, German Cancer Research Center (DKFZ), Heidelberg, Germany.

Clarissa Gerhauser (C)

Division of Epigenomics and Cancer Risk Factors, German Cancer Research Center (DKFZ), Heidelberg, Germany.

Verica Paunović (V)

Institute of Microbiology and Immunology, Faculty of Medicine, University of Belgrade, Belgrade, Serbia.

Anja Tolić (A)

Department of Molecular Biology, Institute for Biological Research "Siniša Stanković" - National Institute of Republic of Serbia, University of Belgrade, Belgrade, Serbia.

Jovana Rajić (J)

Department of Molecular Biology, Institute for Biological Research "Siniša Stanković" - National Institute of Republic of Serbia, University of Belgrade, Belgrade, Serbia.

Svetlana Dinić (S)

Department of Molecular Biology, Institute for Biological Research "Siniša Stanković" - National Institute of Republic of Serbia, University of Belgrade, Belgrade, Serbia.

Aleksandra Uskoković (A)

Department of Molecular Biology, Institute for Biological Research "Siniša Stanković" - National Institute of Republic of Serbia, University of Belgrade, Belgrade, Serbia.

Nevena Grdović (N)

Department of Molecular Biology, Institute for Biological Research "Siniša Stanković" - National Institute of Republic of Serbia, University of Belgrade, Belgrade, Serbia.

Mirjana Mihailović (M)

Department of Molecular Biology, Institute for Biological Research "Siniša Stanković" - National Institute of Republic of Serbia, University of Belgrade, Belgrade, Serbia.

Renata Z Jurkowska (RZ)

School of Biosciences, Cardiff University, Cardiff, Wales, United Kingdom.

Tomasz P Jurkowski (TP)

School of Biosciences, Cardiff University, Cardiff, Wales, United Kingdom.

Jelena Arambašić Jovanović (JA)

Department of Molecular Biology, Institute for Biological Research "Siniša Stanković" - National Institute of Republic of Serbia, University of Belgrade, Belgrade, Serbia.

Melita Vidaković (M)

Department of Molecular Biology, Institute for Biological Research "Siniša Stanković" - National Institute of Republic of Serbia, University of Belgrade, Belgrade, Serbia.

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