title: CRISPR/Cas9-Mediated Genome Editing of Mouse Small Intestinal Organoids.
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.
authors.affiliation: Hubrecht Institute/KNAW, University Medical Center Utrecht, Uppsalalaan 8, Utrecht 3584 CT, The Netherlands. schwankg@ethz.ch.|Institute of Molecular Health Sciences, ETH Zurich, Otto-Stern-Weg 7, HPL, 8093, Zürich, Switzerland. schwankg@ethz.ch.
authors.forename: Gerald
authors.initials: G
authors.lastname: Schwank
authors.affiliation: Hubrecht Institute/KNAW, University Medical Center Utrecht, Uppsalalaan 8, Utrecht 3584 CT, The Netherlands. h.clevers@hubrecht.eu.|Princess Maxima Center for Pediatric Oncology, University Medical Center Utrecht, Uppsalalaan 8, Utrecht 3584 CT, The Netherlands. h.clevers@hubrecht.eu.
authors.forename: Hans
authors.initials: H
authors.lastname: Clevers
mesh_terms: D000818:Animals; D064113:CRISPR-Cas Systems; D000072669:Gene Editing; D007421:Intestine, Small / Q000166:cytology*; D051379:Mice; D009924:Organ Culture Techniques
keywords: CRISPR/Cas9; Genome editing; Intestinal stem cells; Small intestinal organoids