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Nat Commun. 2016 Feb 26;7:10770. doi: 10.1038/ncomms10770.

Multiplexed pancreatic genome engineering and cancer induction by transfection-based CRISPR/Cas9 delivery in mice.

Author information

1
Department of Medicine II, Klinikum rechts der Isar, Technische Universität München, 81675 Munich, Germany.
2
German Cancer Consortium (DKTK), German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany.
3
Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge CB10 1SA, UK.
4
Institute of Radiology, Klinikum rechts der Isar, Technischen Universität München, 81675 Munich, Germany.
5
Department of Pathology, Klinikum rechts der Isar, Technische Universität München, 81675 Munich, Germany.
6
Institute of Virology, Technische Universität München/Helmholtz Zentrum München, 81675 Munich, Germany.
7
Instituto de Biomedicina y Biotecnología de Cantabria, 39011 Santander, Spain.
8
Helmholtz Zentrum München, Research Unit Radiation Cytogenetics, 85764 Neuherberg, Germany.
9
Division of Chronic Inflammation and Cancer, German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany.

Abstract

Mouse transgenesis has provided fundamental insights into pancreatic cancer, but is limited by the long duration of allele/model generation. Here we show transfection-based multiplexed delivery of CRISPR/Cas9 to the pancreas of adult mice, allowing simultaneous editing of multiple gene sets in individual cells. We use the method to induce pancreatic cancer and exploit CRISPR/Cas9 mutational signatures for phylogenetic tracking of metastatic disease. Our results demonstrate that CRISPR/Cas9-multiplexing enables key applications, such as combinatorial gene-network analysis, in vivo synthetic lethality screening and chromosome engineering. Negative-selection screening in the pancreas using multiplexed-CRISPR/Cas9 confirms the vulnerability of pancreatic cells to Brca2-inactivation in a Kras-mutant context. We also demonstrate modelling of chromosomal deletions and targeted somatic engineering of inter-chromosomal translocations, offering multifaceted opportunities to study complex structural variation, a hallmark of pancreatic cancer. The low-frequency mosaic pattern of transfection-based CRISPR/Cas9 delivery faithfully recapitulates the stochastic nature of human tumorigenesis, supporting wide applicability for biological/preclinical research.

PMID:
26916719
PMCID:
PMC4773438
DOI:
10.1038/ncomms10770
[Indexed for MEDLINE]
Free PMC Article
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