Pancreatic cancer modeling using retrograde viral vector delivery and in vivo CRISPR/Cas9-mediated somatic genome editing

Shin Heng Chiou, Ian P. Winters, Jing Wang, Santiago Naranjo, Crissy Dudgeon, Fiona B. Tamburini, Jennifer J. Brady, Dian Yang, Barbara M. Grüner, Chen Hua Chuang, Deborah R. Caswell, Hong Zeng, Pauline Chu, Grace E. Kim, Darren R. Carpizo, Seung K. Kim, Monte M. Winslow

Research output: Contribution to journalArticlepeer-review

114 Scopus citations

Abstract

Pancreatic ductal adenocarcinoma (PDAC) is a genomically diverse, prevalent, and almost invariably fatal malignancy. Although conventional genetically engineered mouse models of human PDAC have been instrumental in understanding pancreatic cancer development, these models are much too labor-intensive, expensive, and slow to perform the extensive molecular analyses needed to adequately understand this disease. Here we demonstrate that retrograde pancreatic ductal injection of either adenoviral-Cre or lentiviral-Cre vectors allows titratable initiation of pancreatic neoplasias that progress into invasive and metastatic PDAC. To enable in vivo CRISPR/Cas9-mediated gene inactivation in the pancreas, we generated a Cre-regulated Cas9 allele and lentiviral vectors that express Cre and a single-guide RNA. CRISPR-mediated targeting of Lkb1 in combination with oncogenic Kras expression led to selection for inactivating genomic alterations, absence of Lkb1 protein, and rapid tumor growth that phenocopied Cremediated genetic deletion of Lkb1. This method will transform our ability to rapidly interrogate gene function during the development of this recalcitrant cancer.

Original languageEnglish (US)
Pages (from-to)1576-1585
Number of pages10
JournalGenes and Development
Volume29
Issue number14
DOIs
StatePublished - Jul 15 2015
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Genetics
  • Developmental Biology

Keywords

  • CRISPR
  • Genome editing
  • Mouse model
  • Pancreatic cancer

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