The CCDC7 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population targeting the human CCDC7 gene. This loss-of-function model generates a heterogeneous pool of HEK293T cells with CRISPR/Cas9-mediated gene disruption, enabling pooled functional genomics and signaling studies without requiring clonal isolation. The product provides a versatile tool for investigating CCDC7??s role in primary cilium assembly and centrosomal biology within an epithelial context.
HEK293T is a human embryonic kidney epithelial cell line stably expressing the SV40 large T antigen, which facilitates episomal plasmid replication and high-efficiency transfection. These cells are widely used for lentivirus production and transient expression assays. Under appropriate conditions, HEK293T cells form primary cilia, making them a suitable model for studying ciliogenesis and related signaling pathways in a well-characterized, robust cell system.
CCDC7 functions as a centriolar satellite protein that interacts with PCM1 and CEP290 to promote primary cilium assembly. Its loss disrupts ciliogenesis, attenuating Hedgehog signaling mediated by the GLI1 and GLI2 transcription factors. Upstream, CCDC7 expression is controlled by RFX transcription factors and cell cycle regulators, while downstream, it influences Hedgehog target genes. Pathway components such as IFT88 further link CCDC7 to intraflagellar transport. Additionally, CCDC7 has been associated with Wnt signaling, positioning it at the intersection of multiple developmental pathways.
In the HEK293T background, CCDC7 knockout provides a reductionist model to dissect the molecular machinery of ciliogenesis and Hedgehog signal transduction. The epithelial origin is relevant for ciliopathy and cancer studies, while the polyclonal population averages clonal variation, mimicking heterogeneous cell pools in screening applications. The robust transfectability of HEK293T allows for complementation and reporter assays, and SV40 T antigen does not impair cilia formation, preserving physiological relevance.
Applications include ciliopathy research, cancer cell signaling, and CRISPR knockout screening. Representative assays are immunofluorescence for acetylated tubulin to assess ciliation, western blotting for GLI1/2, RT?qPCR for GLI target genes, and co-immunoprecipitation with PCM1 for interaction studies. Locus sequencing enables genotype verification. This model supports both mechanistic dissection and high-throughput screens for modulators of ciliogenesis and Hedgehog signaling. For further details, please contact Ascent Research.
