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

作者：Maresch Roman; Mueller Sebastian; Veltkamp Christian; Oellinger 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 Ignacio; Unger Kristian; Yang Fengtang; Schmid Roland M; Vassiliou George S; Braren Rickmer; Schneider Nter; Heikenwalder Mathias; Bradley Allan; Saur Dieter

来源：Nature Communications, 2016, 7(1): 10770.

DOI：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.

• 出版日期2016-2