CRISPR somatic genome engineering and cancer modeling in the mouse pancreas and liver.
Journal
Nature protocols
ISSN: 1750-2799
Titre abrégé: Nat Protoc
Pays: England
ID NLM: 101284307
Informations de publication
Date de publication:
04 2022
04 2022
Historique:
received:
28
07
2021
accepted:
07
12
2021
pubmed:
16
3
2022
medline:
12
4
2022
entrez:
15
3
2022
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 35288718
doi: 10.1038/s41596-021-00677-0
pii: 10.1038/s41596-021-00677-0
doi:
Types de publication
Journal Article
Review
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
1142-1188Subventions
Organisme : Deutsche Forschungsgemeinschaft (German Research Foundation)
ID : DFG RA 1629/2-1
Organisme : Deutsche Forschungsgemeinschaft (German Research Foundation)
ID : SA1374/4-2
Organisme : Deutsche Krebshilfe (German Cancer Aid)
ID : 70114314
Informations de copyright
© 2022. The Author(s), under exclusive licence to Springer Nature Limited.
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