Strategies for genetic manipulation of adult stem cell-derived organoids |
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Authors: | Constantin Menche Henner F Farin |
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Institution: | 1.Georg-Speyer-Haus, Institute for Tumor Biology and Experimental Therapy, Frankfurt am Main, Germany ;2.Frankfurt Cancer Institute, Goethe University, Frankfurt am Main, Germany ;3.German Cancer Consortium (DKTK), Heidelberg, Germany ;4.German Cancer Research Center (DKFZ), Heidelberg, Germany |
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Abstract: | Organoid technology allows the expansion of primary epithelial cells from normal and diseased tissues, providing a unique model for human (patho)biology. In a three-dimensional environment, adult stem cells self-organize and differentiate to gain tissue-specific features. Accessibility to genetic manipulation enables the investigation of the molecular mechanisms underlying cell fate regulation, cell differentiation and cell interactions. In recent years, powerful methodologies using lentiviral transgenesis, CRISPR/Cas9 gene editing, and single-cell readouts have been developed to study gene function and carry out genetic screens in organoids. However, the multicellularity and dynamic nature of stem cell-derived organoids also present challenges for genetic experimentation. In this review, we focus on adult gastrointestinal organoids and summarize the state-of-the-art protocols for successful transgenesis. We provide an outlook on emerging genetic techniques that could further increase the applicability of organoids and enhance the potential of organoid-based techniques to deepen our understanding of gene function in tissue biology.Subject terms: Mechanisms of disease, Genetic engineering, Mutagenesis, Experimental models of disease |
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