CRISPR/Cas9-Mediated Genome Editing of Mouse Small Intestinal Organoids

TY - JOUR

T1 - CRISPR/Cas9-Mediated Genome Editing of Mouse Small Intestinal Organoids

AU - Schwank, Gerald

AU - Clevers, Hans

PY - 2016

Y1 - 2016

N2 - 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.

AB - 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.

U2 - 10.1007/978-1-4939-3603-8_1

DO - 10.1007/978-1-4939-3603-8_1

M3 - Article

C2 - 27246017

SN - 1064-3745

VL - 1422

SP - 3

EP - 11

JO - Methods in Molecular Biology

JF - Methods in Molecular Biology

ER -