Generation of focal mutations and large genomic deletions in the pancreas using inducible in vivo genome editing.

Autor:	Mishra A; Department of Gastroenterology, Hepatology, and Endocrinology, Hannover, Germany., Emamgholi F; Department of Gastroenterology, Hepatology, and Endocrinology, Hannover, Germany., Erlangga Z; Department of Gastroenterology, Hepatology, and Endocrinology, Hannover, Germany., Hartleben B; Department of Pathology, Hannover Medical School, Hannover, Germany., Unger K; Research Unit Radiation Cytogenetics, German Research Center for Environmental Health, Munich, Germany., Wolff K; Department of Gastroenterology, Hepatology, and Endocrinology, Hannover, Germany., Teichmann U; Animal House., Kessel M; Department for Molecular Cell Biology, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany., Woller N; Department of Gastroenterology, Hepatology, and Endocrinology, Hannover, Germany., Kühnel F; Department of Gastroenterology, Hepatology, and Endocrinology, Hannover, Germany., Dow LE; Weill Cornell Graduate School of Medical Sciences, Weill Cornell Medicine, New York, USA., Manns MP; Department of Gastroenterology, Hepatology, and Endocrinology, Hannover, Germany., Vogel A; Department of Gastroenterology, Hepatology, and Endocrinology, Hannover, Germany., Lowe SW; Cancer Biology and Genetics Program, Sloan Kettering Institute, New York, USA., Saborowski A; Department of Gastroenterology, Hepatology, and Endocrinology, Hannover, Germany., Saborowski M; Department of Gastroenterology, Hepatology, and Endocrinology, Hannover, Germany.
Jazyk:	angličtina
Zdroj:	Carcinogenesis [Carcinogenesis] 2020 May 14; Vol. 41 (3), pp. 334-344.
DOI:	10.1093/carcin/bgz108
Abstrakt:	Beyond the nearly uniform presence of KRAS mutations, pancreatic cancer is increasingly recognized as a heterogeneous disease. Preclinical in vivo model systems exist, but with the advent of precision oncology, murine models with enhanced genetic flexibility are needed to functionally annotate genetic alterations found in the human malignancy. Here, we describe the generation of focal gene disruptions and large chromosomal deletions via inducible and pancreas-specific expression of Cas9 in adult mice. Experimental mice are derived on demand directly from genetically engineered embryonic stem cells, without the need for further intercrossing. To provide initial validation of our approach, we show that disruption of the E3 ubiquitin ligase Rnf43 accelerates KrasG12D-dependent tumourigenesis. Moreover, we demonstrate that this system can be used to rapidly interrogate the impact of complex cancer-associated alleles through the generation of a previously unstudied 1.2 megabase deletion surrounding the CDKN2A and CDKN2B tumour suppressors. Thus, our approach is capable of reproducibly generating biallelic and precise loss of large chromosomal fragments that, in conjunction with mutant Kras, leads to development of pancreatic ductal adenocarcinoma with full penetrance. (© The Author(s) 2019. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)
Databáze:	MEDLINE
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