The Generation of Organoids for Studying Wnt Signaling

Jarno Drost, Benedetta Artegiani, Hans Clevers

Research output: Contribution to journal/periodicalArticleScientificpeer-review

Abstract

We established an in vitro culture model in which intestinal epithelial stem cells can grow into three-dimensional, ever-expanding epithelial organoids that retain their original organ identity and genetic stability. Moreover, organoids can easily be genetically modified using different genome modification strategies, including viral delivery of transgenes and CRISPR/Cas9 technology. These combined characteristics make them a useful in vitro model system to study many biological processes including the contribution of cellular signaling pathways to tissue homeostasis and disease. Here we describe our current laboratory protocols to establish human intestinal organoids and how to genetically modify both mouse and human intestinal organoids to study cellular signaling pathways, specifically Wnt signaling. Moreover, we provide a detailed protocol for lentiviral transduction and CRISPR/Cas9-mediated genome modification of organoid cultures.

Original languageEnglish
Pages (from-to)141-59
Number of pages19
JournalMethods in Molecular Biology
Volume1481
DOIs
Publication statusPublished - 2016

Keywords

  • Journal Article

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