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Multiplexed pancreatic genome engineering and cancer induction by transfection-based CRISPR/Cas9 delivery in mice.
- Source :
-
Nature communications [Nat Commun] 2016 Feb 26; Vol. 7, pp. 10770. Date of Electronic Publication: 2016 Feb 26. - Publication Year :
- 2016
-
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.
- Subjects :
- Animals
BRCA2 Protein genetics
CRISPR-Cas Systems
Chromosome Deletion
Electroporation
Genetic Engineering methods
Genome
High-Throughput Nucleotide Sequencing
Immunohistochemistry
Magnetic Resonance Imaging
Mice
Mutation
Neoplasms, Experimental genetics
Phylogeny
Polymerase Chain Reaction
Proto-Oncogene Proteins p21(ras) genetics
Sequence Analysis, DNA
Transfection methods
Translocation, Genetic genetics
Carcinogenesis genetics
Pancreas metabolism
Pancreatic Neoplasms genetics
Subjects
Details
- Language :
- English
- ISSN :
- 2041-1723
- Volume :
- 7
- Database :
- MEDLINE
- Journal :
- Nature communications
- Publication Type :
- Academic Journal
- Accession number :
- 26916719
- Full Text :
- https://doi.org/10.1038/ncomms10770