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Nat Commun. 2015 Jun 11;6:7391. doi: 10.1038/ncomms8391.

Somatic CRISPR/Cas9-mediated tumour suppressor disruption enables versatile brain tumour modelling.

Author information

1
Division of Molecular Genetics, German Cancer Research Center (DKFZ), Im Neuenheimer Feld 280, 69120 Heidelberg, Germany.
2
1] Department of Neuropathology, Heinrich Heine University Düsseldorf, Moorenstrasse 5, 40225 Düsseldorf, Germany [2] German Cancer Consortium (DKTK), partner site Essen/Düsseldorf, 40225 Düsseldorf, Germany.
3
Division of Pediatric Neurooncology, German Cancer Research Center (DKFZ), Im Neuenheimer Feld 280, 69120 Heidelberg, Germany.
4
Masonic Cancer Center, University of Minnesota, Minneapolis, Minnesota 55455, USA.
5
Tumor Cell Biology, St Jude Children's Research Hospital, 262 Danny Thomas Place, Memphis, Tennessee 38105, USA.
6
1] Department of Neuropathology, University of Heidelberg, Im Neuenheimer Feld 220, 69120 Heidelberg, Germany [2] Clinical Cooperation Unit Neuropathology, German Cancer Research Center (DKFZ), Im Neuenheimer Feld 220-221, 69120 Heidelberg, Germany.

Abstract

In vivo functional investigation of oncogenes using somatic gene transfer has been successfully exploited to validate their role in tumorigenesis. For tumour suppressor genes this has proven more challenging due to technical aspects. To provide a flexible and effective method for investigating somatic loss-of-function alterations and their influence on tumorigenesis, we have established CRISPR/Cas9-mediated somatic gene disruption, allowing for in vivo targeting of TSGs. Here we demonstrate the utility of this approach by deleting single (Ptch1) or multiple genes (Trp53, Pten, Nf1) in the mouse brain, resulting in the development of medulloblastoma and glioblastoma, respectively. Using whole-genome sequencing (WGS) we characterized the medulloblastoma-driving Ptch1 deletions in detail and show that no off-targets were detected in these tumours. This method provides a fast and convenient system for validating the emerging wealth of novel candidate tumour suppressor genes and the generation of faithful animal models of human cancer.

PMID:
26067104
PMCID:
PMC4467376
DOI:
10.1038/ncomms8391
[Indexed for MEDLINE]
Free PMC Article

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