Multiplexed pancreatic genome engineering and cancer induction by transfection-based CRISPR/Cas9 delivery in mice.
Schmid, Roland M
Vassiliou, George S
Springer Science and Business Media LLC
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Maresch, R., Mueller, S., Veltkamp, C., Öllinger, R., Friedrich, M., Heid, I., Steiger, K., et al. (2016). Multiplexed pancreatic genome engineering and cancer induction by transfection-based CRISPR/Cas9 delivery in mice.. Nat Commun, 7 10770. https://doi.org/10.1038/ncomms10770
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.
Pancreas, Animals, Mice, Pancreatic Neoplasms, Neoplasms, Experimental, Chromosome Deletion, Translocation, Genetic, BRCA2 Protein, Magnetic Resonance Imaging, Electroporation, Immunohistochemistry, Transfection, Genetic Engineering, Polymerase Chain Reaction, Sequence Analysis, DNA, Phylogeny, Mutation, Genome, Proto-Oncogene Proteins p21(ras), High-Throughput Nucleotide Sequencing, Carcinogenesis, CRISPR-Cas Systems
Medical Research Council (MC_PC_12009)
External DOI: https://doi.org/10.1038/ncomms10770
This record's URL: https://www.repository.cam.ac.uk/handle/1810/311495
Attribution 4.0 International
Licence URL: https://creativecommons.org/licenses/by/4.0/