The CC2D1A Knockout 786-O Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population in which target-gene disruption has been performed on the endogenous CC2D1A locus in the human 786-O cell line. This polyclonal format yields a heterogeneous pool of gene-edited cells, enabling researchers to study loss-of-function phenotypes while avoiding potential artifacts arising from single-cell cloning. The population is suitable for directly establishing functional models of CC2D1A deficiency in a relevant cancer cell background.
The parental 786-O line was derived from a primary clear cell renal cell adenocarcinoma and is a widely accepted model of ccRCC. These cells harbor a well-characterized VHL tumor suppressor mutation, leading to constitutive stabilization of the hypoxia-inducible factor HIF-1??. Consequently, 786-O cells exhibit dysregulated hypoxia signaling, elevated pro-angiogenic factor expression, and aberrant transcriptional programs that underlie key aspects of renal carcinoma progression.
CC2D1A encodes a scaffold protein that functions as a critical negative regulator of NF-??B signaling. Mechanistically, it binds directly to the RELA (p65) subunit of NF-??B and represses its transcriptional activity toward target genes such as IL6, TNF, and BCL2L1. Moreover, CC2D1A interacts with the ESCRT-III component CHMP4B to modulate endosomal sorting, linking membrane trafficking to transcriptional control. Upstream, its activity is regulated by the IKK complex (IKK??, IKK??, and IKK??/NEMO) and by calcium signaling via its C2 domain, while it also associates with histone deacetylase HDAC1 to exert transcriptional repression.
In the VHL-mutant, HIF-1??-expressing 786-O background, disruption of CC2D1A removes a major restraint on NF-??B activation, resulting in enhanced basal and stimulus-responsive NF-??B pathway activity. This creates a powerful cellular model to dissect how CC2D1A normally counterbalances the pro-inflammatory and pro-survival gene expression often driven by VHL loss in ccRCC. Furthermore, altered CHMP4B-mediated endosomal trafficking may impact receptor turnover and signaling dynamics, providing additional insights into tumor cell adaptation and drug resistance.
These polyclonal knockout cells are optimized for mechanistic studies of NF-??B signaling, endosomal trafficking, and renal carcinoma biology. Typical experimental approaches include NF-??B luciferase reporter assays to quantify pathway activation, RT-qPCR profiling of downstream target genes (e.g., IL6, TNF, BCL2L1), and co-immunoprecipitation to validate interactions with RELA, CHMP4B, or IKBKB. The model supports phenotypic analyses such as cell proliferation, migration, and drug resistance assays. For additional technical specifications or customization options, please contact Ascent Research.
