| Title: |
Multiplexed pancreatic genome engineering and cancer induction by transfection-based CRISPR/Cas9 delivery in mice |
| Authors: |
Maresch, Roman; Mueller, Sebastian; Veltkamp, Christian; Ollinger, Rupert; Friedrich, Mathias; Heid, Irina; Steiger, Katja; Weber, Julia; Engleitner, Thomas; Barenboim, Maxim; Klein, Sabine; Louzada, Sandra; Banerjee, Ruby; Strong, Alexander; Stauber, Teresa; Gross, Nina; Geumann, Ulf; Lange, Sebastian; Ringelhan, Marc; Varela Egocheaga, Ignacio |
| Contributors: |
Universidad de Cantabria |
| Source: |
Nature Communications 7, Article number: 10770 (2016) |
| Publisher Information: |
Nature Publishing Group |
| Publication Year: |
2016 |
| Collection: |
Universidad de Cantabria: UCrea |
| Description: |
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. |
| Document Type: |
article in journal/newspaper |
| Language: |
English |
| Relation: |
https://hdl.handle.net/10902/10989 |
| DOI: |
10.1038/ncomms10770 |
| Availability: |
https://hdl.handle.net/10902/10989; https://doi.org/10.1038/ncomms10770 |
| Rights: |
This work is licensed under a Creative Commons Attribution 4.0 International License. ; openAccess |
| Accession Number: |
edsbas.9588F4F2 |
| Database: |
BASE |