TY  - JOUR
AU  - Kaltenbacher, Thorsten
AU  - Löprich, Jessica
AU  - Maresch, Roman
AU  - Weber, Julia
AU  - Müller, Sebastian
AU  - Oellinger, Rupert
AU  - Groß, Nina
AU  - Griger, Joscha
AU  - de Andrade Krätzig, Niklas
AU  - Avramopoulos, Petros
AU  - Ramanujam, Deepak
AU  - Brummer, Sabine
AU  - Widholz, Sebastian A
AU  - Bärthel, Stefanie
AU  - Falcomatà, Chiara
AU  - Pfaus, Anja
AU  - Alnatsha, Ahmed
AU  - Mayerle, Julia
AU  - Schmidt-Supprian, Marc
AU  - Reichert, Maximilian
AU  - Schneider, Günter
AU  - Ehmer, Ursula
AU  - Braun, Christian Jörg
AU  - Saur, Dieter Karl Maximilian
AU  - Engelhardt, Stefan
AU  - Rad, Roland
TI  - CRISPR somatic genome engineering and cancer modeling in the mouse pancreas and liver.
JO  - Nature protocols
VL  - 17
IS  - 4
SN  - 1750-2799
CY  - Basingstoke
PB  - Nature Publishing Group
M1  - DKFZ-2022-00488
SP  - 1142-1188
PY  - 2022
N1  - 2022 Apr;17(4):1142-1188
AB  - 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.
LB  - PUB:(DE-HGF)16
C6  - pmid:35288718
DO  - DOI:10.1038/s41596-021-00677-0
UR  - https://inrepo02.dkfz.de/record/179131
ER  -