The CCDC185 Knockout HEK293T Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal population of HEK293T cells with targeted disruption of the CCDC185 gene, providing a robust loss-of-function model for investigating centrosomal biology and ciliogenesis. This polyclonal pool avoids clonal isolation bottlenecks and maintains population-level phenotypic diversity while effectively silencing CCDC185.
HEK293T is an immortalized human embryonic kidney epithelial cell line expressing SV40 large T antigen, renowned for high transfection efficiency and robust protein expression. Its epithelial origin permits the study of centrosomal and ciliary processes, as these cells can form primary cilia under serum-starvation conditions, making them a suitable host for interrogating CCDC185 function.
CCDC185 is a centrosomal coiled-coil domain protein essential for centriole assembly and primary cilium formation. It is transcriptionally regulated by E2F and FOXM1 and activated by CDK1 and Aurora A kinases. CCDC185 interacts with CEP135, CEP250, pericentrin, and ???tubulin to organize the centrosome and mitotic spindle. Downstream, it promotes ciliogenesis and Hedgehog signaling, increasing GLI1 and GLI2 expression, while also modulating cell cycle regulators Cyclin B1 and CDK1. The pathway involves PLK4, CEP152, SAS?6, STIL, and CPAP, positioning CCDC185 at a critical node for centrosome duplication and ciliary signaling.
In HEK293T cells, CCDC185 knockout likely disrupts centriole architecture and blocks ciliogenesis, attenuating Hedgehog signal transduction. This defect may alter cell cycle progression and mitotic fidelity, as centrosomal aberrations can activate spindle checkpoints. The polyclonal knockout population thus offers a relevant model to examine how loss of a key centrosomal protein impacts epithelial cell homeostasis and signaling.
Applications include centrosome and ciliogenesis research, Hedgehog pathway studies, and ciliopathy drug screening. Typical assays involve immunofluorescence for ???tubulin and pericentrin, cilium formation assays with acetylated tubulin, Western blotting for GLI1 and ARL13B, cell cycle flow cytometry, and proliferation assays (MTT, EdU). This polyclonal knockout cell product is a versatile tool for dissecting centrosome-related pathologies. For more information, contact Ascent Research.
