The CCDC85B Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population in which the CCDC85B gene has been disrupted in the human HAP1 near-haploid cell line. This heterogeneous pool of knockout cells serves as a loss-of-function model for investigating the cellular roles of the coiled-coil domain-containing protein CCDC85B, which is implicated in centrosome function and ciliogenesis. The polyclonal format provides a robust tool for functional genomics studies, enabling assessment of gene disruption effects across a mixed genetic background without clonal selection artifacts.
HAP1 cells are a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia (CML) model, characterized by a stable haploid karyotype that simplifies genetic manipulation and phenotypic analysis. Originating from leukemic cells, they retain key signaling and proliferative properties relevant to cancer biology, while their haploid state facilitates straightforward knockout generation and reduces genetic redundancy. This makes HAP1 an ideal host for CRISPR/Cas9-mediated gene disruption, especially for studying essential genes in pathways that may exhibit lethality in diploid cells.
CCDC85B encodes a centrosome-localized coiled-coil domain protein that functions in cilia assembly and cell cycle progression. It is regulated upstream by cell cycle-dependent kinases (CDKs) and the transcription factor E2F1, linking its expression to proliferative control. Mechanistically, CCDC85B interacts with centrosomal protein CEP170, tubulin, and its paralog CCDC85A to influence ciliary transport and microtubule organization. Disruption of CCDC85B is predicted to impair ciliary signaling, potentially affecting Hedgehog pathway components such as the receptor SMO and transcription factor GLI2, as well as cyclin A/CDK1 complexes that drive cell cycle transitions.
In the HAP1 cellular context, CCDC85B knockout may unmask phenotypic consequences related to centrosome duplication and primary cilium formation, processes frequently dysregulated in cancer and ciliopathies. The near-haploid background allows for efficient screening of these phenotypes with minimal confounding from wild-type alleles, making the model particularly valuable for dissecting the gene??s role in cell cycle regulation and developmental signaling. Loss of CCDC85B could also impair Hedgehog-dependent transcriptional programs, offering insights into its potential contributions to tumorigenesis and developmental defects.
The CCDC85B Knockout HAP1 Polyclonal Cells are suitable for a range of experimental applications, including immunofluorescence microscopy to assess cilia markers, flow cytometry-based cell cycle analysis, western blotting for cell cycle proteins, RT-qPCR profiling of Hedgehog pathway genes, and proliferation assays to evaluate growth defects. This knockout model is a versatile resource for centrosome biology, ciliogenesis research, and drug target validation in a haploid human system. For additional technical information, please contact Ascent Research.
