The EDIL3 Knockout 786-O Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population generated from the 786-O human clear cell renal cell carcinoma (ccRCC) cell line. This product provides a loss-of-function model for the EDIL3 gene, enabling investigation of its roles in tumor biology and vascular signaling. The polyclonal pool contains a heterogeneous mixture of EDIL3-disrupted cells, offering a robust tool for functional studies without the clonal selection bias associated with monoclonal lines. CRISPR/Cas9-mediated gene disruption was used to target EDIL3, resulting in a population of cells with ablated EDIL3 expression, suitable for examining EDIL3-dependent phenotypes in a cancer-relevant context.
The parental 786-O cell line is a widely used model of ccRCC, derived from a primary renal cell adenocarcinoma of a male patient. These epithelial cells harbor a mutated von Hippel-Lindau (VHL) tumor suppressor gene, leading to constitutive stabilization of hypoxia-inducible factors (HIFs), particularly HIF-1??, even under normoxic conditions. This VHL-deficient background mimics the genetic hallmark of the majority of clear cell RCCs, making 786-O cells a physiologically relevant platform for studying pathways dysregulated in renal cancer, including angiogenesis, metabolic reprogramming, and cell adhesion.
EDIL3 (EGF-like repeats and discoidin I-like domains 3) is a secreted extracellular matrix protein that functions as a ligand for integrin receptors ??v??3 and ??v??5. Upon binding, EDIL3 activates focal adhesion kinase (FAK) and Src kinase, triggering downstream signaling cascades including the PI3K/AKT and ERK/MAPK pathways. These pathways promote endothelial cell adhesion, migration, and survival, while simultaneously inhibiting NF-??B-mediated leukocyte adhesion and inflammatory responses. In the tumor microenvironment, EDIL3 is transcriptionally upregulated by HIF-1?? in response to hypoxia, and its expression is further modulated by inflammatory cytokines such as TNF-?? and IL-1??. EDIL3 also interacts with phosphatidylserine and other ECM components, integrating signals that regulate vascular development and remodeling. By disrupting EDIL3, the knockout cells allow dissection of integrin-mediated signaling independently of other ECM factors.
In the context of VHL-deficient 786-O cells, EDIL3 is a key effector of the pseudohypoxic state driven by HIF-1?? stabilization. The loss of VHL leads to elevated EDIL3 expression, which contributes to the angiogenic phenotype and metastatic potential of ccRCC. EDIL3 knockout in these cells provides a direct system to interrogate tumor-intrinsic and paracrine effects on endothelial cell behavior, such as tube formation and migration in co-culture with HUVECs. This model is particularly valuable for elucidating the crosstalk between tumor cells and the vascular niche, and for assessing the role of EDIL3 in mediating resistance to anti-angiogenic therapies like sunitinib. The polyclonal nature of the knockout pool ensures that the observed phenotypes are not artifacts of clonal variation, strengthening the reproducibility of functional assays.
Researchers can employ the EDIL3 Knockout 786-O Polyclonal Cells in a variety of assay formats. Western blotting and RT-qPCR confirm EDIL3 ablation, while migration and invasion assays evaluate metastatic potential. Phospho-AKT and phospho-ERK analysis probes downstream signaling. Co-culture with HUVECs enables tube formation assays to quantify angiogenesis. Cell adhesion and flow cytometry experiments assess integrin surface levels. Drug sensitivity testing with sunitinib or other targeted agents can reveal EDIL3-dependent therapeutic responses. Additionally, these cells are suitable for genetic screens or for examining interactions with immune cells in the inflammatory milieu. For further information or to discuss custom applications, please contact Ascent Research.
