The CCDC97 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the human CCDC97 gene in the HEK293T host cell line. This heterogeneous pool of knockout cells, generated via CRISPR/Cas9-mediated gene disruption, enables robust loss-of-function studies without single-cell cloning. The polyclonal format reduces clonal variation and simplifies population-level phenotypic analyses. These cells are ideal for investigating CCDC97-dependent processes in a high-transfectability human embryonic kidney model.
The HEK293T cell line is an immortalized human embryonic kidney epithelial line expressing SV40 large T antigen, which enables high transfection efficiency and recombinant protein production. This host is a workhorse for biochemical and screening applications, with adherent growth and flat morphology suitable for imaging. Its epithelial origin provides a relevant context for studying microtubule organization and ciliary biology, as primary cilia are present in many epithelial cells.
CCDC97 encodes a coiled-coil domain protein predicted to stabilize microtubules and facilitate ciliogenesis. It interacts with tubulin, and likely functions with other coiled-coil proteins and microtubule-associated proteins to regulate cytoskeletal dynamics. Disruption of CCDC97 may alter microtubule stability, impair cilium formation, and affect cilium-dependent signaling. This knockout model enables direct investigation of CCDC97??s role in microtubule and ciliary processes.
In HEK293T cells, CCDC97 disruption may produce phenotypes related to microtubule network integrity and primary cilium assembly. These cells can form primary cilia under serum starvation, allowing assessment of ciliary length and composition by immunofluorescence. Microtubule dynamics also influence migration and cell cycle progression; thus, CCDC97 knockout may affect these processes. The polyclonal pool averages stochastic effects, yielding a stable loss-of-function population for reproducible quantitative assays. Rescue constructs or fluorescent reporters can be transfected to dissect CCDC97 function with high throughput.
Applications include live-cell imaging of GFP-tagged tubulin for microtubule dynamics, ciliary biology assays with acetylated ??-tubulin and Arl13b immunostaining, and RNA-seq transcriptomic profiling. Migration assays and cell cycle analysis by flow cytometry can further characterize phenotypes. RT-qPCR and Western blotting confirm knockout and quantify downstream changes. These cells are suitable for high-content screening for ciliopathy phenotypes or microtubule modulators. For additional details or to obtain this knockout model, please contact Ascent Research.
