The CCDC138 Knockout HEK293T Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the widely utilized HEK293T host cell line. This product provides a heterogeneous pool of cells harboring targeted disruptions in the CCDC138 gene, enabling robust loss-of-function studies without clonal selection. The polyclonal format preserves genetic diversity and mitigates clonal artifacts, making it suitable for population-level analyses of gene function. These cells are designed for researchers investigating ciliary biology and microtubule-associated processes, offering a versatile platform for phenotypic and biochemical assays.
HEK293T cells are a human embryonic kidney cell line transformed with adenovirus type 5 DNA and constitutively expressing the SV40 large T antigen, which enhances episomal replication of plasmids containing the SV40 origin of replication. They are extensively employed for high-titer viral production, transient and stable protein expression, and signal transduction studies. While HEK293T cells do not typically form primary cilia under standard adherent culture conditions, they can be induced to undergo ciliogenesis upon serum starvation or specific differentiation protocols, thereby enabling the investigation of ciliary assembly and disassembly mechanisms. This host background provides a tractable system for ectopic expression and functional complementation experiments.
CCDC138 (coiled-coil domain-containing protein 138), also known as CFAP126, is predicted to be a cilia- and flagella-associated protein that functions as a structural component of the axonemal microtubule scaffold. Mechanistically, CCDC138 is believed to contribute to microtubule stability and dynein-mediated motility, potentially regulated by the master ciliogenic transcription factor FOXJ1 and RFX-family transcription factors, including RFX3. It likely interacts directly with microtubules and axonemal dynein arm complexes, and is implicated in ciliogenesis, axonemal dynein assembly, and spermatogenesis. Representative pathway components include FOXJ1, RFX3, DNAI1, and DNAH5, with CCDC138 acting downstream of transcriptional activation and upstream of axonemal dynein function.
In the HEK293T context, disruption of CCDC138 provides a model to dissect the role of this protein in early ciliary assembly steps, particularly under conditions that stimulate ciliogenesis. Even in the absence of full cilia formation, CCDC138 knockout may impair microtubule organization or intracellular trafficking pathways that are relevant to the cell’s known roles in viral packaging and protein expression. This model thus enables the study of CCDC138-dependent processes in a well-characterized, easily transfectable background, facilitating structure-function analyses and the mapping of protein interaction networks. The polyclonal nature of the knockout population also allows assessment of heterogeneity in ciliogenic responses.
Researchers can employ these cells for a variety of experimental applications, including functional characterization of ciliary proteins, investigation of microtubule-associated processes, and disease modeling related to ciliopathies and potential infertility phenotypes. Representative assays include Western blotting to confirm knockout efficiency and assess downstream targets, immunofluorescence microscopy for ciliary markers such as acetylated alpha-tubulin and ARL13B, co-immunoprecipitation to identify interacting partners, RT-qPCR to quantify ciliary gene expression changes, and ciliogenesis induction assays with serum starvation. For additional information or technical support, please contact Ascent Research.
