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

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 Cre-mediated genetic deletion of Lkb1. This method will transform our ability to rapidly interrogate gene function during the development of this recalcitrant cancer.

Modeling invasive lobular breast carcinoma by CRISPR/Cas9-mediated somatic genome editing of the mammary gland

“…For example, liver-specific gene disruption was achieved by transient delivery of components of the CRISPR/Cas9 system in the tail veins of mice, leading to hepatocellular carcinoma Weber et al 2015). Similar approaches have been used to deliver targeted oncogenic mutations to the lung (Platt et al 2014;Sánchez-Rivera et al 2014), brain (Zuckermann et al 2015), and pancreas (Chiou et al 2015).…”

Ethics Dumping

“…A recent example has been the creation of cancer models in mice, where the cancerous mutation was introduced through genome editing using viral vectors -in essence transforming cancer into a transmissible infectious disease (Chiou et al 2015). This creates novel safety risks that will need to be included in biosafety oversight schemes.…”

Ips Progression a Decade Devoted

“…Successful progression of iPS and CRISPR-CAS amalgamation is justified by stable iPS proliferation and wholesome numbers of modified clones that can be harvested after genetic modification in culture. Immunodeficiency centromeric region instability, facial anomalies syndrome (ICF) and pancreatic cancer models have been generated using the CRISPR-CAS editing system [95,96]. Seamless gene correction of β-thalassemia mutations in patient-specific iPSCs has been possible using CRISPR/ Cas9 system [97,98].…”