Abstract
The CRISPR/Cas9 system is an RNA-guided genome-editing tool that has been recently developed based on the bacterial CRISPR-Cas immune defense system. Due to its versatility and simplicity, it rapidly became the method of choice for genome editing in various biological systems, including mammalian cells. Here we describe a protocol for CRISPR/Cas9-mediated genome editing in murine small intestinal organoids, a culture system in which somatic stem cells are maintained by self-renewal, while giving rise to all major cell types of the intestinal epithelium. This protocol allows the study of gene function in intestinal epithelial homeostasis and pathophysiology and can be extended to epithelial organoids derived from other internal mouse and human organs.
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Acknowledgments
This work was funded by grants from the European Research Council (EU/232814-StemCeLLMark), the KNAW/3V-fund, the SNF (31003A_160230), and the Human Frontiers in Science Program long-term fellowship LT000422/2012.
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Schwank, G., Clevers, H. (2016). CRISPR/Cas9-Mediated Genome Editing of Mouse Small Intestinal Organoids. In: Ivanov, A. (eds) Gastrointestinal Physiology and Diseases. Methods in Molecular Biology, vol 1422. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3603-8_1
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DOI: https://doi.org/10.1007/978-1-4939-3603-8_1
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3601-4
Online ISBN: 978-1-4939-3603-8
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