CRISPR somatic genome engineering and cancer modeling in the mouse pancreas and liver

Genetically engineered mouse models (GEMMs) transformed the study of organismal disease phenotypes but are limited by their lengthy generation in embryonic stem cells. Here, we describe methods for rapid and scalable genome engineering in somatic cells of the liver and pancreas through delivery of C...

Full description

Saved in:

Bibliographic Details
Published in:	Nature protocols Vol. 17; no. 4; pp. 1142 - 1188
Main Authors:	Kaltenbacher, Thorsten, Löprich, Jessica, Maresch, Roman, Weber, Julia, Müller, Sebastian, Oellinger, Rupert, Groß, Nina, Griger, Joscha, de Andrade Krätzig, Niklas, Avramopoulos, Petros, Ramanujam, Deepak, Brummer, Sabine, Widholz, Sebastian A., Bärthel, Stefanie, Falcomatà, Chiara, Pfaus, Anja, Alnatsha, Ahmed, Mayerle, Julia, Schmidt-Supprian, Marc, Reichert, Maximilian, Schneider, Günter, Ehmer, Ursula, Braun, Christian J., Saur, Dieter, Engelhardt, Stefan, Rad, Roland
Format:	Journal Article
Language:	English
Published:	London Nature Publishing Group UK 01-04-2022 Nature Publishing Group
Subjects:	631/1647/1511 631/1647/767/70 631/208/4041/3196 631/67/1504/1610 631/67/1504/1713 Analytical Chemistry Animal models Animals Biological Techniques Biomedical and Life Sciences Cancer Chromosomes Clustered Regularly Interspaced Short Palindromic Repeats - genetics Computational Biology/Bioinformatics CRISPR CRISPR-Cas Systems - genetics Data analysis Design of experiments Embryo cells Experimental design Gene Editing - methods Genetic engineering Genetic modification Genome editing Genomes Hepatocytes Life Sciences Liver Mice Mice, Knockout Microarrays Modelling Multiplexing Neoplasms - genetics Organic Chemistry Pancreas Phenotypes Protocol Somatic cells Stem cell transplantation Stem cells
Online Access:	Get full text
Tags:	Add Tag No Tags, Be the first to tag this record!

Description
Summary:	Genetically engineered mouse models (GEMMs) transformed the study of organismal disease phenotypes but are limited by their lengthy generation in embryonic stem cells. Here, we describe methods for rapid and scalable genome engineering in somatic cells of the liver and pancreas through delivery of CRISPR components into living mice. We introduce the spectrum of genetic tools, delineate viral and nonviral CRISPR delivery strategies and describe a series of applications, ranging from gene editing and cancer modeling to chromosome engineering or CRISPR multiplexing and its spatio-temporal control. Beyond experimental design and execution, the protocol describes quantification of genetic and functional editing outcomes, including sequencing approaches, data analysis and interpretation. Compared to traditional knockout mice, somatic GEMMs face an increased risk for mouse-to-mouse variability because of the higher experimental demands of the procedures. The robust protocols described here will help unleash the full potential of somatic genome manipulation. Depending on the delivery method and envisaged application, the protocol takes 3–5 weeks. The authors provide protocols for rapid and scalable genome engineering in somatic cells of the liver and pancreas through both viral and nonviral delivery of CRISPR components into living mice.
Bibliography:	ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-3 content type line 23 ObjectType-Review-1
ISSN:	1754-2189 1750-2799
DOI:	10.1038/s41596-021-00677-0