Npc1l1 Knockout Panc02 Cell Line

The Npc1l1 Knockout Panc02 Cell Line is a CRISPR/Cas9-edited murine knockout cell line generated from the Panc02 pancreatic ductal adenocarcinoma parental line. This product provides a stable loss-of-function model for the Npc1l1 gene, which encodes the Niemann-Pick C1-like 1 (NPC1L1) cholesterol transporter. Through CRISPR/Cas9-mediated target-gene disruption, researchers can ablate NPC1L1 expression to investigate its role in cholesterol metabolism and pancreatic cancer biology. The cell line serves as a defined genetic model for studying sterol transport mechanisms and for preclinical evaluation of NPC1L1-targeted therapeutics.

The host Panc02 cell line is a chemically induced pancreatic adenocarcinoma derived from C57BL/6 mice. It is a well-established model of pancreatic cancer, exhibiting tumorigenic properties in syngeneic hosts. Panc02 cells are widely used to study pancreatic cancer progression, metastasis, and metabolism, as well as to test immunotherapeutic strategies. Their murine origin allows for in vivo implantation studies in immunocompetent C57BL/6 recipients, facilitating translational research.

NPC1L1 is a polytopic transmembrane protein critical for the apical uptake of dietary and biliary cholesterol in enterocytes and hepatocytes. Its expression is transcriptionally regulated by SREBP2, LXR, and HNF4?? in response to cellular sterol levels. Upon cholesterol binding, NPC1L1 interacts with flotillin-1 and flotillin-2, recruits the AP2 complex and clathrin, and undergoes Rab protein-mediated endocytosis. Internalized cholesterol is delivered to the endoplasmic reticulum for esterification and incorporation into chylomicrons or hepatic VLDL particles. NPC1L1 functions within a network that includes the heterodimeric sterol exporters ABCG5/ABCG8 and modulates downstream effects on bile acid synthesis via CYP7A1 and LDLR expression, thereby influencing systemic cholesterol homeostasis.

In pancreatic ductal adenocarcinoma, dysregulated cholesterol metabolism supports membrane biogenesis, signaling platform formation, and proliferation. The Panc02 knockout model enables dissection of NPC1L1-dependent cholesterol uptake in the tumor microenvironment. Because NPC1L1 is the molecular target of the cholesterol-lowering drug ezetimibe, this cell line allows for target engagement studies and assessment of ezetimibe-mediated effects on cancer cell growth, lipid droplet accumulation, and lipoprotein secretion. It also provides a platform to explore cross-talk between sterol transport and oncogenic pathways in a pancreatic cancer context.

Typical applications include cholesterol uptake assays using radiolabeled or fluorescent cholesterol, filipin staining to visualize free cholesterol distribution, and western blot analysis of NPC1L1 and associated proteins. Genotyping by PCR and Sanger sequencing of the targeted locus confirm gene disruption. The knockout line can be employed in proliferation and viability assays, ezetimibe treatment experiments to validate target specificity, and lipoprotein secretion profiling. It is also suitable for co-culture studies investigating lipid-mediated signaling within the tumor microenvironment. For additional product details, protocols, or technical support, please contact Ascent Research.