The KCTD9 Knockout 786-O Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the 786-O human renal cell carcinoma line. This loss-of-function model, created via CRISPR/Cas9-mediated disruption of KCTD9, enables investigation of KCTD9 function in a tumorigenic kidney epithelial background. The polyclonal format reduces clonal bias and is suitable for studies of ubiquitin-mediated signaling, NF-??B regulation, and apoptotic control.
The 786-O cell line, established from a primary clear cell renal cell carcinoma, harbors a VHL mutation leading to constitutive HIF-1?? stabilization. This VHL-deficient, HIF-1??-overexpressing phenotype drives oncogenic processes such as angiogenesis and survival signaling, making 786-O cells a key model for RCC research. The epithelial origin supports studies of polarity, migration, and invasion in a cancer context.
KCTD9 encodes a substrate adaptor for the Cullin3-RING E3 ubiquitin ligase (CRL3) complex, interacting with Cullin3 and RBX1 to ubiquitinate I??B??, thereby promoting its degradation and NF-??B p65 activation. This mechanism positions KCTD9 as a positive regulator of NF-??B signaling, triggered by TNF-?? via the TNFR?CIKK??/IKK?? axis. KCTD9 also modulates apoptosis and is itself regulated by NF-??B and miR-181a, forming feedback loops. Its role in protein ubiquitination links it to signal termination and stress responses.
In 786-O cells, KCTD9 knockout is expected to disrupt I??B?? ubiquitination, leading to I??B?? stabilization and attenuated NF-??B activity. Given the high HIF-1??-driven stress, KCTD9 loss may sensitize cells to apoptosis or alter chemosensitivity. This model facilitates dissection of how CRL3KCTD9-mediated NF-??B regulation integrates with VHL/HIF signaling to influence tumor cell fitness, drug resistance, and invasive behavior.
These polyclonal knockout cells can be used in Western blotting for I??B?? and phospho-p65, NF-??B luciferase reporter assays, co-immunoprecipitation of the CRL3 complex, and ubiquitination studies. Functional assays such as Annexin V/PI apoptosis profiling, scratch wound migration, and transwell invasion can assess phenotypic consequences. Drug sensitivity screens (e.g., cisplatin) are also applicable. This product is ideal for probing KCTD9-dependent signaling, NF-??B pathway inhibitor screening, and exploring vulnerabilities in VHL-deficient RCC. For ordering or technical assistance, contact Ascent Research.
