The CCDC12 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population designed to disrupt the endogenous CCDC12 gene in HEK293T cells. This product comprises a pool of cells with heterogeneous editing outcomes at the CCDC12 locus, collectively yielding a loss-of-function model for the centrosomal coiled-coil domain protein. The polyclonal format captures a spectrum of editing outcomes, avoiding clonal bias and enabling robust assessment of CCDC12-dependent phenotypes in a physiologically relevant context.
HEK293T cells are a commonly used human embryonic kidney epithelial line expressing SV40 large T-antigen, enabling episomal plasmid replication. Their adherent, epithelial-like morphology and high transfection efficiency make them ideal for recombinant protein expression, lentivirus production, and gene editing. These cells retain fundamental aspects of epithelial cell biology, including centrosome and primary cilium regulation, facilitating studies on ciliogenesis and cell division.
CCDC12 encodes a centrosomal coiled-coil domain protein that localizes to the pericentriolar material and is essential for centrosome integrity and microtubule organization. Its activity is regulated by CDK1/cyclin B and PLK1 phosphorylation during the cell cycle. CCDC12 interacts with centriolar and centrosomal components, including CEP135, CEP152, and PCM1, and is implicated in centriole duplication and spindle assembly. Downstream, CCDC12 influences microtubule nucleation and ciliary axoneme assembly, linking it to both mitotic spindle formation and ciliogenesis. CRISPR/Cas9-mediated disruption of CCDC12 perturbs these processes, leading to defective spindle morphology and impaired cilia formation.
In HEK293T cells, CCDC12 knockout provides a tractable model for centrosome biology and ciliopathy research. HEK293T cells do not constitutively form primary cilia but can be induced by serum starvation, enabling convenient ciliogenesis assays. The polyclonal knockout population exhibits a range of functional perturbations, allowing correlation of editing outcomes with cellular phenotypes. This model recapitulates centrosome dysfunction features observed in human diseases, such as spindle assembly defects and ciliogenesis failure, and supports genetic and chemical modifier screens.
This polyclonal CCDC12 knockout product is suited for a variety of experimental applications, including immunofluorescence microscopy for centrosome markers, cilia formation assays, cell cycle analysis by flow cytometry, and co-immunoprecipitation studies of centrosomal interactions. Western blotting confirms CCDC12 depletion. The polyclonal format permits investigation of loss-of-function heterogeneity. For additional information, please contact Ascent Research.
