The CCN2 Knockout 786-O Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from the 786-O human renal clear cell adenocarcinoma epithelial cell line, featuring targeted disruption of the CCN2 gene. This polyclonal knockout pool provides a heterogeneous loss-of-function model for investigating connective tissue growth factor (CTGF) biology in kidney cancer and fibrosis research.
The 786-O parental line originates from a primary clear cell renal carcinoma and is widely utilized as an in vitro model of renal cell carcinoma (RCC). These cells harbor a homozygous VHL frameshift mutation and carry wild-type PTEN, resulting in constitutive stabilization of hypoxia-inducible factors (HIF-1??/2??) and recapitulating the most common genetic alteration in sporadic clear cell RCC. As adherent epithelial cells, they offer a robust platform for dissecting oncogenic and fibrogenic signaling networks.
CCN2 (CTGF) is a secreted matricellular protein that regulates cell adhesion, migration, proliferation, extracellular matrix (ECM) synthesis, and angiogenesis. It functions as a canonical downstream mediator of transforming growth factor-beta (TGF-??) signaling. Activation of TGFBR1/2 by TGFB1 leads to phosphorylation of SMAD2 and SMAD3, which complex with SMAD4 to drive CCN2 transcription. Extracellular CTGF interacts with integrins ??V??3 and ??5??1, LRP1, and heparan sulfate proteoglycans, triggering FAK and SRC kinase activation. This cascade promotes expression of ECM components (COL1A1, COL1A2, fibronectin) and matrix metalloproteinases (MMP2, MMP9), while upregulating EMT transcription factors SNAI1 and TWIST1. CCN2 expression is also induced by hypoxia, mechanical stress, and angiotensin II, linking TGF-??, HIF, and mechanotransduction pathways.
In the VHL-deficient 786-O background, CCN2 knockout is expected to attenuate TGF-??-driven ECM deposition, integrin signaling, and EMT-related gene expression, potentially reducing cell migration and invasion. This polyclonal knockout population enables dissection of CTGF-dependent phenotypes in renal carcinoma without clonal bias, preserving the diversity of CRISPR editing outcomes for robust comparative studies.
These cells are suited for fibrosis research, renal cell carcinoma tumor progression studies, drug target validation, and anti-fibrotic drug screening. Key assays include TGF-??1-induced collagen gel contraction, Boyden chamber migration and invasion, RT-qPCR for CCN2 and downstream targets, western blotting for CTGF and phospho-SMAD2/3, immunofluorescence for E-cadherin/vimentin, and luciferase reporter assays for SMAD-responsive elements. For further technical details or custom inquiries, please contact Ascent Research.
