Format

Send to

Choose Destination
Carcinogenesis. 2019 Jun 6. pii: bgz108. doi: 10.1093/carcin/bgz108. [Epub ahead of print]

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

Author information

1
Department of Gastroenterology, Hepatology, and Endocrinology, Hannover Medical School, Hannover, Lower Saxony, Germany.
2
Department of Pathology, Hannover Medical School, Hannover, Lower Saxony, Germany.
3
Research Unit Radiation Cytogenetics, German Research Center for Environmental Health, Munich, Bavaria, Germany.
4
Animal House, Max Planck Institute for Biophysical Chemistry, Göttingen, Lower Saxony, Germany.
5
Department for Molecular Cell Biology, Max Planck Institute for Biophysical Chemistry, Göttingen, Lower Saxony, Germany.
6
Weill Cornell Graduate School of Medical Sciences, Weill Cornell Medicine, New York, New York, USA.
7
Cancer Biology and Genetics Progam, Sloan Kettering Institute, New York, New York, USA.

Abstract

Beyond the nearly uniform presence of KRAS mutations, pancreatic cancer is increasingly recognized as a heterogeneous disease. Pre-clinical 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 CKDN2A 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.

PMID:
31170286
DOI:
10.1093/carcin/bgz108

Supplemental Content

Full text links

Icon for Silverchair Information Systems
Loading ...
Support Center