The C2CD3 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population targeting the C2CD3 gene. This engineered pool of HAP1 cells carries heterogeneous gene disruptions, providing a loss-of-function model that avoids single-clone artefacts while enabling robust investigation of C2CD3-dependent processes. The polyclonal format preserves genetic diversity and is ideally suited for population-level functional genomics, pooled screening, and biochemical studies where clonal variability is undesirable. Researchers can exploit this model to dissect the cellular consequences of C2CD3 deficiency without the confounding effects of monoclonal selection.
The host HAP1 cell line is a near-haploid human cell line originally derived from KBM-7 chronic myeloid leukemia cells. Its near-haploid karyotype simplifies genetic manipulation and facilitates the generation of clear loss-of-function phenotypes, as most loci are present in a single copy. HAP1 cells retain expression of key ciliary and signaling components, making them a versatile and robust platform for studying pathways that intersect with centrosome biology and ciliogenesis. Their rapid proliferation and ease of culture further enhance their utility for high-throughput imaging, biochemical time courses, and CRISPR-based functional screens.
C2CD3 encodes a distal centriolar protein that is essential for centriole elongation and the initiation of ciliogenesis. It functions as a positive regulator of ciliary vesicle formation by recruiting OFD1, CEP120, and CPAP to the distal end of the mother centriole, thereby promoting assembly of the distal appendage and subsequent axoneme extension. C2CD3 also interacts with TALPID3, a centrosomal protein implicated in centriole maturation. The protein localizes downstream of Hedgehog signal transduction and is transcriptionally regulated by ciliogenic factors such as RFX3 and FOXJ1. In the absence of C2CD3, centriole elongation fails, ciliary vesicle docking is impaired, and Hedgehog signaling is disrupted, mirroring key pathogenic events in ciliopathies.
In the HAP1 near-haploid context, C2CD3 knockout creates a simplified genetic environment for dissecting the molecular hierarchy of centriole assembly and ciliary signaling. Because HAP1 cells rely on the same core centriolar machinery as other human cells, the knockout phenotype provides a tractable model for structure?Cfunction studies of distal centriole components. The loss-of-function background enables unambiguous assignment of C2CD3-dependent interactions and downstream signaling events. Moreover, the haploid nature may unmask recessive cellular defects that are masked in diploid models, offering a powerful complement to studies in primary cells or organoids.
This knockout model is particularly valuable for disease-modeling applications related to orofaciodigital syndrome type 14, Joubert syndrome, and broader centrosome-related ciliopathies. Experimental applications include immunofluorescence analysis of centriolar markers (e.g., CEP120, CPAP), ciliogenesis assays under serum-starvation conditions, western blotting for C2CD3 and its interaction partners, RT-qPCR profiling of ciliary gene expression, and Hedgehog reporter assays to quantify pathway activity. These cells are suitable for drug discovery screens targeting ciliogenesis or Hedgehog signaling, as well as for mechanistic studies of distal centriole biology. For further details or technical support, please contact Ascent Research.
