The CCDC50 Knockout HEK293T Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal population with disruption of the CCDC50 gene. Unlike monoclonal knockouts, this mixed pool avoids clonal artifacts while retaining the genetic heterogeneity necessary for robust loss-of-function studies. The knockout was achieved through CRISPR/Cas9-mediated gene disruption, and the product is supplied as a population of cells with reduced CCDC50 expression. This format is well-suited for experiments requiring bulk assessment of signaling pathways without single-cell clone limitations.
HEK293T cells are human embryonic kidney epithelial cells that stably express the SV40 large T-antigen, enabling high-efficiency transient transfection and viral production. Derived from HEK293 cells, they maintain endogenous expression of key signaling components, including TNFR1, EGFR, NEDD4, and TRAF2, making them an ideal platform for studying NF-??B, JNK, and ubiquitin-dependent pathways. Their well-characterized genome and ease of manipulation have established HEK293T as a standard host for gene perturbation experiments in signal transduction and cancer biology.
CCDC50 is a cytosolic adaptor protein that regulates TNF-??-induced NF-??B activation and EGFR degradation. Through its interaction with NEDD4 and TRAF2, CCDC50 promotes ubiquitination and lysosomal targeting of EGFR, thereby attenuating downstream signaling. It also modulates the IKK??/I??B??/NF-??B axis, controlling transcription of pro-inflammatory genes such as IL-6 and TNF. Additionally, CCDC50 influences JNK/AP-1 signaling, placing it at the intersection of cell survival, inflammation, and apoptosis. Its ability to interact with ESCRT components further highlights its role in endosomal sorting and receptor turnover.
In HEK293T cells, knockout of CCDC50 allows dissection of its molecular functions in a relevant epithelial context. The polyclonal nature of the population enables evaluation of signaling changes without clonal bias, revealing how loss of CCDC50 alters NF-??B reporter activity, EGFR degradation kinetics, and apoptotic thresholds. Because HEK293T endogenously express TRAF2, NEDD4, and EGFR, the cellular model supports native interaction studies and avoids the artifacts associated with overexpression systems. This makes it particularly useful for studying ubiquitin-mediated regulation of receptor tyrosine kinases.
Typical applications include NF-??B luciferase reporter assays, phospho-I??B?? immunoblotting, and EGFR degradation time-course experiments. Co-immunoprecipitation of NEDD4 or TRAF2 can be employed to probe disrupted protein complexes, while RT-qPCR for IL-6 and TNF quantifies transcriptional outputs. Apoptosis assays using Annexin V further enable assessment of cell death signaling. These tools position the CCDC50 Knockout HEK293T Polyclonal Cells as a versatile model for investigating inflammatory diseases, cancer signaling, and the molecular basis of DFNA44 hearing loss. For further details, please contact Ascent Research.
