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

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Stem Cells and Lineage Commitment

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2736))

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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.

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Acknowledgments

The authors thank Max Pietschmann for expert advice and support on establishing the silencing method. The authors also acknowledge research funding received from the Research Council of Norway (NFR 251041 and 314397), Novo Nordic Foundation (NNF15OC0015054 and NNF21OC0067325), and the Norwegian Diabetesforbundet forskningfond.

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Authors and Affiliations

  1. Mohn Research Center for Diabetes Precision Medicine, Department of Clinical Science, Faculty of Medicine, University of Bergen, Bergen, Norway

    Lucas Unger, Luiza Ghila & Simona Chera

Authors
  1. Lucas Unger
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  2. Luiza Ghila
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  3. Simona Chera
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Correspondence to Simona Chera .

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Editors and Affiliations

  1. Ottawa, ON, Canada

    Kursad Turksen

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Unger, L., Ghila, L., Chera, S. (2023). Targeted Gene Silencing by Using GapmeRs in Differentiating Human-Induced Pluripotent Stem Cells (hiPSC) Toward Pancreatic Progenitors. In: Turksen, K. (eds) Stem Cells and Lineage Commitment. Methods in Molecular Biology, vol 2736. Humana, New York, NY. https://doi.org/10.1007/7651_2023_498

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  • DOI: https://doi.org/10.1007/7651_2023_498

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3536-0

  • Online ISBN: 978-1-0716-3537-7

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