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Maresch, R.* ; Müller, S.* ; Veltkamp, C.* ; Öllinger, R.* ; Friedrich, M.* ; Heid, I.M.* ; Steiger, K.* ; Weber, J.* ; Engleitner, T.* ; Barenboim, M.* ; Klein, S.* ; Louzada, S.* ; Banerjee, R.* ; Strong, A.* ; Stauber, T.* ; Gross, N.* ; Geumann, U.* ; Lange, S.* ; Ringelhan, M. ; Varela, I.* ; Unger, K. ; Yang, F.* ; Schmid, R.M.* ; Vassiliou, G.S.* ; Braren, R.* ; Schneider, G.* ; Heikenwälder, M. ; Bradley, A.* ; Saur, D.* ; Rad, R.*

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

Nat. Commun. 7:10770 (2016)
Publ. Version/Full Text Supplement Research data DOI
Open Access Gold
Creative Commons Lizenzvertrag
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.
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Publication type Article: Journal article
Document type Scientific Article
Keywords In-vivo; Gene-transfer; Chromosomal Rearrangements; Ductal Adenocarcinoma; Immune-system; Mouse Models; Plasmid Dna; Cell-lines; Stem-cells; Instability
ISSN (print) / ISBN 2041-1723
e-ISSN 2041-1723
Quellenangaben Volume: 7, Issue: , Pages: , Article Number: 10770 Supplement: ,
Publisher Nature Publishing Group
Publishing Place London
Reviewing status Peer reviewed