Targeted Gene Silencing by Using GapmeRs in Differentiating Human-Induced Pluripotent Stem Cells (hiPSC) Toward Pancreatic Progenitors

Lucas Unger; Luiza Ghila; Simona Chera

doi:10.1007/7651_2023_498

Targeted Gene Silencing by Using GapmeRs in Differentiating Human-Induced Pluripotent Stem Cells (hiPSC) Toward Pancreatic Progenitors

Methods Mol Biol. 2024:2736:23-38. doi: 10.1007/7651_2023_498.

Authors

Lucas Unger¹, Luiza Ghila¹, Simona Chera²

Affiliations

¹ Mohn Research Center for Diabetes Precision Medicine, Department of Clinical Science, Faculty of Medicine, University of Bergen, Bergen, Norway.
² Mohn Research Center for Diabetes Precision Medicine, Department of Clinical Science, Faculty of Medicine, University of Bergen, Bergen, Norway. Simona.Chera@uib.no.

PMID: 37615889
DOI: 10.1007/7651_2023_498

Abstract

Induced pluripotent stem cells as a source for generating pancreatic islet endocrine cells represent a great research tool for deciphering the molecular mechanisms of lineage commitment, a layered multi-step process. Additionally, targeted gene silencing by using GapmeRs, short antisense oligonucleotides, proved instrumental in studying the role of different developmental genes. Here we describe our approach to induce mTOR silencing by using specific GapmeRs during the differentiation of induced pluripotent stem cells toward pancreatic progenitors. We will describe our current differentiation protocol, the transfection procedure, and the quality control steps required for a successful experiment.

Keywords: GapmeR; Gene silencing; Human iPSC; In vitro differentiation; Islet endocrine cells; Pancreatic endocrine progenitors.

Publication types

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

MeSH terms

Cell Differentiation
Gene Silencing
Humans
Induced Pluripotent Stem Cells*
Pancreas
Transfection