The CCDC85C Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the near-haploid human HAP1 cell line, with disruption of the CCDC85C gene. This format provides a genetically diverse loss-of-function pool suitable for robust functional studies without the need for clonal isolation.
HAP1 cells originate from the KBM-7 chronic myeloid leukemia line and maintain a near-haploid karyotype, which simplifies genetic manipulation and phenotypic interpretation. Their fibroblast-like morphology and stable growth make them ideal for high-resolution imaging and genetic screens. Widely used in functional genomics, HAP1 cells serve as a model for leukemia and haploid genetic studies.
CCDC85C is a centriolar satellite protein that regulates primary cilium formation and centriole duplication by facilitating protein trafficking to the centrosome. It interacts with CEP164, CEP290, and PCM1 within the centriolar satellite network. Upstream, RFX transcription factors and Wnt signaling regulate its expression, while downstream it influences Hedgehog pathway activity via GLI1 and GLI2 transcription factors. Key pathway components include IFT particles, the BBSome, Hedgehog ligands and receptors, and Aurora kinase A. Loss of CCDC85C impairs ciliogenesis and disrupts Hedgehog signaling, contributing to ciliopathies such as Joubert syndrome, Meckel syndrome, and nephronophthisis.
The near-haploid HAP1 background reduces genetic redundancy, enhancing the phenotypic effects of CCDC85C disruption. This model is valuable for dissecting centriolar satellite biology and Hedgehog signaling, and for screening ciliopathy-related drugs. It also allows investigation of centrosome abnormalities in a leukemia context, bridging cilia biology and oncogenic signaling.
These polyclonal knockout cells are suited for serum starvation-induced ciliogenesis assays with immunofluorescence detection of Arl13b and acetylated tubulin, Western blotting for centriolar proteins, and RT-qPCR analysis of Hedgehog targets like GLI1 and PTCH1. Additional applications include flow cytometry for cell cycle analysis and high-throughput screening for Hedgehog pathway modulators. For more information, contact Ascent Research.
