CRISPR Somatic Genome Engineering and Cancer Modeling in the Mouse Pancreas and Liver

Overview

Journal Nat Protoc

Specialties Biology
Pathology
Science

Date 2022 Mar 15

PMID 35288718

Authors

Thorsten Kaltenbacher

Jessica Loprich

Roman Maresch

Julia Weber

Sebastian Muller

Rupert Oellinger

Nina Gross

Joscha Griger

Niklas de Andrade Kratzig

Petros Avramopoulos

Deepak Ramanujam

Sabine Brummer

Sebastian A Widholz

Stefanie Barthel

Chiara Falcomata

Anja Pfaus

Ahmed Alnatsha

Julia Mayerle

Marc Schmidt-Supprian

Maximilian Reichert

Gunter Schneider

Ursula Ehmer

Christian J Braun

Dieter Saur

Stefan Engelhardt

Roland Rad

Affiliations

Soon will be listed here.

Abstract

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.

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