The CCDC89 Knockout HAP1 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population designed for dissection of centrosome biology and cell cycle regulation. This non-clonal population carries targeted disruption of the CCDC89 gene within the near-haploid HAP1 cell line, producing a heterogeneous loss-of-function model that retains the genetic accessibility of the host background while enabling pooled screening and population-level phenotypic analyses. The polyclonal format avoids clonal artifacts and supports robust assessment of CCDC89-dependent processes across a spectrum of editing events.
The HAP1 host cell line is a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia (CML) isolate from a 40-year-old male. These cells are BCR-ABL positive and have a predominantly haploid karyotype, making them exceptionally well-suited for genome-wide knockout screens, CRISPR-based functional genomics, and gene-trap experiments. The near-haploid genome simplifies mutational analysis and reduces genetic redundancy, while the CML origin provides a relevant context for studying oncogenic signaling and centriole-dependent cell division mechanisms.
CCDC89 encodes a centrosomal protein essential for centriole duplication and microtubule organization. Mechanistically, CCDC89 functions within the canonical centriole biogenesis pathway, acting downstream of PLK4 and CEP152 and interacting directly with CEP63, CEP135, and CDK5RAP2 to promote procentriole assembly. Its activity is regulated by the E2F1 transcriptional program and cell cycle kinases including CDK2-cyclin E. Disruption of CCDC89 perturbs recruitment of downstream centriolar proteins such as CEP152, CEP63, and CDK5RAP2, and impairs proper nucleation by the gamma-tubulin ring complex. These molecular connections place CCDC89 at a convergence point for signals that govern centrosome duplication, mitotic spindle organization, and primary ciliogenesis.
In the HAP1 background, ablation of CCDC89 creates a valuable model for exploring centrosome dysfunction. The near-haploid state amplifies the phenotypic consequences of gene disruption, yielding pronounced centriole duplication defects, mitotic errors, and G1/S cell cycle arrest. These outcomes are directly relevant to centrosome-related disorders such as microcephaly and ciliopathies, as well as cancer research where aberrant centrosome number and mitotic fidelity contribute to genomic instability. The polyclonal knockout pool allows simultaneous examination of multiple loss-of-function variants, facilitating dose-response studies and genetic modifier screens that are not achievable with single clones.
This product supports a broad range of applications in centrosome and cell cycle research. Typical assays include immunofluorescence detection of centrosomal markers (??-tubulin, CPAP) to quantify centriole numbers, Western blotting to confirm CCDC89 depletion, RT-qPCR for transcriptional profiling, and flow cytometry for DNA content analysis. Centrosome duplication assays and EdU proliferation measurements enable functional assessment of centriole biogenesis, while the knockout cells can be employed in drug sensitivity testing of anti-mitotic compounds. For additional information or custom inquiries, please contact Ascent Research.
