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

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...

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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
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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. 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.
AbstractList 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.
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.
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.
Author Müller, Sebastian
Engelhardt, Stefan
Braun, Christian J.
Saur, Dieter
Schneider, Günter
Löprich, Jessica
Oellinger, Rupert
Bärthel, Stefanie
Falcomatà, Chiara
Pfaus, Anja
Mayerle, Julia
Groß, Nina
Avramopoulos, Petros
Widholz, Sebastian A.
Ramanujam, Deepak
Kaltenbacher, Thorsten
Brummer, Sabine
Alnatsha, Ahmed
de Andrade Krätzig, Niklas
Ehmer, Ursula
Reichert, Maximilian
Schmidt-Supprian, Marc
Rad, Roland
Weber, Julia
Griger, Joscha
Maresch, Roman
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BackLink https://www.ncbi.nlm.nih.gov/pubmed/35288718$$D View this record in MEDLINE/PubMed
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ContentType Journal Article
Copyright The Author(s), under exclusive licence to Springer Nature Limited 2022
2022. The Author(s), under exclusive licence to Springer Nature Limited.
The Author(s), under exclusive licence to Springer Nature Limited 2022.
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Snippet Genetically engineered mouse models (GEMMs) transformed the study of organismal disease phenotypes but are limited by their lengthy generation in embryonic...
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SubjectTerms 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
Title CRISPR somatic genome engineering and cancer modeling in the mouse pancreas and liver
URI https://link.springer.com/article/10.1038/s41596-021-00677-0
https://www.ncbi.nlm.nih.gov/pubmed/35288718
https://www.proquest.com/docview/2647964055
https://search.proquest.com/docview/2639223964
Volume 17
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