The CCDC14 Knockout HAP1 Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal knockout cell population derived from the near-haploid human HAP1 cell line. This product provides a loss-of-function model for the CCDC14 gene, which encodes a scaffold protein essential for centriole duplication and mitotic spindle organization. The polyclonal format reflects a pool of edited cells with disruptive mutations at the target locus, enabling functional studies without clonal selection artifacts. The near-haploid background further simplifies genotype?Cphenotype correlations by reducing functional redundancy often observed in diploid models.
The host HAP1 cell line is a near-haploid, suspension-adapted cell line derived from the chronic myelogenous leukemia (CML) KBM-7 line. Its predominantly haploid karyotype (with a disomic region on chromosome 8) allows efficient CRISPR/Cas9-mediated gene disruption, as a single targeting event can produce a functional knockout. This genetic simplicity, combined with robust growth in suspension culture and a stable male phenotype, supports reproducible high-throughput and imaging-based workflows, making it an ideal platform for studying fundamental cellular processes.
CCDC14 functions as a molecular scaffold that bridges the CEP63?CCEP152 complex to downstream centriole assembly factors, acting downstream of PLK4 kinase and CDK2/Cyclin E activity to license centriole duplication. It interacts directly with CEP63 and CEP152, and its loss disrupts the recruitment of essential centriole biogenesis proteins including SAS-6, STIL, and CPAP, thereby blocking procentriole formation. Additional associations with PCM1 and NEDD1 link CCDC14 to pericentriolar material organization and microtubule nucleation. Consequently, CCDC14 knockout leads to defective centriole duplication, aberrant mitotic spindle assembly, and G2/M checkpoint activation, consistent with its role in maintaining centrosome number and genomic stability.
In the HAP1 near-haploid context, CCDC14 disruption yields a clear phenotype characterized by centrosome amplification, multipolar spindle formation, and mitotic delay. These cellular defects mirror the pathological features of primary microcephaly, Seckel syndrome, and centrosome amplification-driven malignancies. The polyclonal knockout population preserves the diversity of editing events, better recapitulating the spectrum of loss-of-function mutations encountered in disease than clonal isolates. This makes the model particularly valuable for genetic interaction mapping, synthetic lethality screens, and drug sensitivity profiling in cancers with underlying centrosome abnormalities.
Key research applications include quantification of centriole numbers via immunofluorescence for centrin or CEP135, centrosome duplication assays using synchronization protocols, and western blot analysis of pathway components such as phospho-PLK4, SAS-6, or STIL. Co-immunoprecipitation experiments can assess the integrity of the CEP63/CEP152 complex, while flow cytometry and mitotic morphology scoring provide complementary readouts of cell cycle distribution and spindle defects. For additional technical information or support, please contact Ascent Research.
