The CCDC50 Knockout HT29 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma cell line, engineered to disrupt the CCDC50 gene. This polyclonal knockout product provides a heterogeneous loss-of-function model that conserves the genetic diversity inherent to pooled editing events, enabling robust functional studies of CCDC50 in a physiologically relevant intestinal epithelial context.
The parental HT29 cell line is an extensively characterized model of human colorectal cancer, displaying epithelial morphology and harboring mutations in the APC and TP53 tumor suppressor genes while retaining wild-type KRAS. Classified within the CMS3 metabolic subtype, HT29 cells are widely employed as an in vitro system for intestinal epithelial biology, particularly for dissecting signaling pathways that govern inflammation-driven colorectal carcinogenesis.
CCDC50 encodes a coiled-coil domain-containing adaptor protein that functions as a negative regulator of NF-??B signaling and a modulator of autophagy. Mechanistically, CCDC50 is activated by pro-inflammatory stimuli such as TNF-??, IL-1??, and TLR ligands, and it acts upstream of the NF-??B pathway by recruiting the deubiquitinase A20 (TNFAIP3) to the IKK complex scaffold protein NEMO (IKBKG), thereby removing activating ubiquitin chains and suppressing I??B?? phosphorylation and p65 nuclear translocation. Additionally, CCDC50 interacts with p62/SQSTM1 to influence autophagic flux, as evidenced by altered LC3-II turnover. Representative pathway components include TRAF6, the IKK complex, and the autophagy protein ATG5.
In the HT29 cellular environment, disruption of CCDC50 removes a critical brake on NF-??B activation, leading to sustained inflammatory signaling that is particularly relevant to colorectal cancer pathophysiology. Given that HT29 cells exhibit constitutive Wnt pathway activation due to APC mutation, the enhanced NF-??B activity upon CCDC50 loss may synergize with oncogenic drivers to promote tumor cell survival, proliferation, and cytokine production. Moreover, the parallel deregulation of autophagy expands the model’s utility for studying the interplay between nutrient-sensing, inflammation, and tumor progression.
This polyclonal knockout population is ideally suited for a broad array of research applications, including the dissection of NF-??B transcriptional responses, autophagy regulation, and inflammation?Ccancer crosstalk in a colorectal epithelial background. Representative assays include Western blotting for phospho-I??B?? and nuclear p65 to monitor pathway activation, NF-??B luciferase reporter assays to quantify transcriptional output, LC3 turnover measurements to assess autophagic flux, co-immunoprecipitation of CCDC50 binding partners such as NEMO or p62, and multiplex cytokine secretion profiling. These cells also serve as a potent tool for drug sensitivity screening against compounds targeting inflammatory or autophagic pathways. For additional technical information, please contact Ascent Research.
