The CCDC116 Knockout HEK293T Polyclonal Cells product provides a CRISPR/Cas9-mediated polyclonal knockout cell population targeting the CCDC116 gene in the human HEK293T cell line. This mixed population contains cells with diverse gene disruption events, offering a versatile loss-of-function model for investigating CCDC116’s oncogenic functions without the limitations of single-clone selection. By employing this population, researchers can study averaged phenotypic effects and pathway modulation relevant to cancer biology.
The host cell model, HEK293T, is a widely used derivative of human embryonic kidney HEK293 cells that stably expresses the SV40 large T antigen. This genetic modification allows for high-copy-number plasmid replication and significantly enhanced transfection efficiency, making the cell line a preferred system for protein expression, viral packaging, and cell signaling studies. The adherent epithelial morphology and robust growth characteristics further contribute to its utility in high-throughput screening and microscopy-based assays.
CCDC116 is a coiled-coil domain-containing oncoprotein that plays a pivotal role in promoting cell proliferation and invasion. It is activated by upstream signals including EGF/EGFR, MYC, and E2F transcription factors, and in turn stimulates the MAPK/ERK cascade??through components such as RAS, BRAF, MEK1/2, and ERK1/2??and the PI3K/AKT pathway via AKT1 and mTOR. Downstream, CCDC116 upregulates Cyclin D1 to drive cell cycle progression and enhances the expression of matrix metalloproteinases MMP2 and MMP9 to facilitate extracellular matrix degradation. Mechanistically, CCDC116 interacts with cytoskeletal proteins (actin, tubulin) and kinases (PAK1, ROCK1) to orchestrate actin cytoskeleton reorganization, thereby promoting cell motility.
In the HEK293T background, these signaling networks are well-represented, enabling precise dissection of CCDC116 function. The SV40 large T antigen binds and inactivates p53 and Rb, which can interplay with oncogenic pathways; however, the robust expression of MAPK/ERK and PI3K/AKT components makes this model ideal for studying CCDC116-mediated proliferative and migratory phenotypes. The polyclonal knockout format minimizes clonal variability and artifacts, allowing evaluation of gene disruption across a heterogeneous population??particularly advantageous for drug target validation and screening campaigns where consistency is critical.
This product is designed for a broad range of research applications, including functional characterization of CCDC116 in cancer biology, investigation of signal transduction mechanisms, and screening for small-molecule inhibitors. Representative assays such as Western blotting, RT-qPCR, and RNA-seq enable transcript and protein profiling; immunofluorescence and flow cytometry facilitate spatial and quantitative analysis; transwell migration/invasion assays, wound healing, and soft agar colony formation measure metastatic and tumorigenic potential; co-immunoprecipitation and phospho-kinase arrays elucidate protein interactions and pathway activation. This model is particularly relevant for colorectal and gastric cancer research. For technical inquiries, please contact Ascent Research.
