CCDC14 Knockout HEK293T Polyclonal Cells constitute a CRISPR/Cas9-mediated gene-disrupted polyclonal population in which the CCDC14 gene is targeted across a heterogeneous pool of HEK293T cells. This product delivers a versatile loss-of-function model that allows researchers to examine the collective consequences of CCDC14 ablation without clonal isolation, thus preserving population-level diversity while eliminating target gene function. The polyclonal format is particularly suited for pooled screening strategies, population-based complementation experiments, and functional studies where batch-to-batch consistency of a defined knockout pool is advantageous over single-cell clones.
Derived from the extensively characterized HEK293T cell line, these cells originate from human embryonic kidney epithelium and are immortalized through stable expression of the SV40 large T antigen. This antigenic background enables high-copy episomal replication of plasmids containing an SV40 origin, making HEK293T cells an industry-standard host for high-level transient protein expression, lentiviral packaging, and receptor co-expression assays. The robust transfectability and rapid growth kinetics of HEK293T provide an optimal cellular chassis for CRISPR-edited knockout models, ensuring reliable delivery of editing components and consistent maintenance of the disrupted genotype under standard culture conditions.
At the molecular level, CCDC14 encodes a centrosomal coiled-coil domain protein that localizes to the pericentriolar matrix and acts as a critical scaffold for PLK1 activation during the G2/M transition. CCDC14 interacts directly with CEP164 and CEP290, core components of the centriole and distal appendages, and facilitates the recruitment and phosphorylation of PLK1 by upstream kinases including Aurora A and CDK1?Ccyclin B. This activation step is essential for downstream events such as centrosome maturation, spindle pole organization, and the initiation of primary cilium assembly. Disruption of CCDC14 thereby uncouples PLK1 from its spatial regulators, leading to impaired activation of the PLK1?CCDC25C?Ccyclin B positive-feedback loop and defective ciliary recruitment of IFT machinery components, including IFT88 and IFT20, as well as tubulin and Gli transcription factors.
In the HEK293T host context, CCDC14 knockout provides a uniquely powerful platform for dissecting centrosome-to-cilium signaling independent of oncogenic transformation artifacts common in many tumor lines. The epithelial origin of HEK293T cells retains core ciliogenesis competency, enabling researchers to study the dual role of CCDC14 in cell cycle progression and cilia formation within a single experimentally tractable system. This model recapitulates the centrosomal dysregulation seen in ciliopathies such as Meckel syndrome type 1 and Joubert syndrome, where mutations in CCDC14-interacting proteins lead to overlapping phenotypes. The high transfectability further allows for re-expression of wild-type or disease-mutant CCDC14 constructs, facilitating structure?Cfunction analyses and the identification of functional domains required for PLK1 binding and cilia initiation.
Research applications of this knockout model span ciliogenesis assays under serum-starvation conditions, immunofluorescence-based visualization of centrosomal and ciliary markers, and quantitative biochemical analyses of PLK1 phosphorylation status by western blotting. Flow cytometric cell cycle profiling can reveal G2/M arrest phenotypes, while co-immunoprecipitation experiments using the HAUS complex or CEP164 enable mapping of protein?Cprotein interaction networks disrupted by CCDC14 loss. The model is also suited for proliferation and viability screens aimed at identifying synthetic lethal interactions or small-molecule modulators of centrosome function, with direct relevance to ciliopathy drug discovery. For additional information or technical support, please contact Ascent Research.
