The CCDC93 Knockout HAP1 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt target gene expression of CCDC93 in the HAP1 human near-haploid cell line. This loss-of-function model enables researchers to investigate the cellular functions of CCDC93 without reliance on pharmacological inhibition or RNA interference, providing a stable genetic background for functional studies. The polyclonal nature of the knockout pool ensures representation of diverse editing events, offering a practical tool for screening experiments and pathway analysis.
The HAP1 cell line is a male-derived human near-haploid chronic myeloid leukemia line originating from the KBM-7 lineage. Its near-haploid karyotype simplifies genetic manipulation and facilitates knockout generation, as only one allele needs to be disrupted to achieve a functional null phenotype in many loci. HAP1 cells are widely employed as a haploid genetics model and a versatile functional genomics screening platform, enabling high-throughput interrogation of gene function in a human context.
CCDC93 encodes an essential scaffolding component of the WASH (Wiskott-Aldrich Syndrome Protein and SCAR Homolog) complex regulatory module. This protein directly interacts with multiple WASH complex members, including WASHC1, WASHC2, WASHC3, WASHC4, and WASHC5, as well as accessory factors such as KIAA1033 and C16orf62. CCDC93 cooperates with the retromer components VPS35 and VPS29 to spatially organize branched actin assembly on endosomal membranes through activation of the Arp2/3 complex. Downstream, CCDC93-dependent regulation promotes F-actin polymerization and endosomal tubulation, which are critical for the efficient sorting and recycling of internalized transmembrane cargo proteins, notably integrin beta-1 (ITGB1) and transferrin receptor (TFRC). While upstream regulatory signals controlling CCDC93 remain uncharacterized, its central role in the WASH complex positions it as a key node linking endosomal membrane dynamics to the actin cytoskeleton.
Disruption of CCDC93 in the HAP1 background is anticipated to impair WASH complex-mediated actin polymerization on endosomes, leading to defects in retromer-dependent cargo sorting and endosomal tubule formation. This knockout model therefore provides a physiologically relevant system to dissect the mechanistic interplay between the WASH complex and retromer in governing endosomal recycling pathways. The near-haploid nature of HAP1 cells further streamlines genetic complementation studies and enables clean genotype-phenotype correlations in trafficking assays, offering an advantage over diploid cell models where compensatory mechanisms may mask knockout effects.
The CCDC93 Knockout HAP1 Polyclonal Cells are suited for a diverse array of experimental applications, including functional genomics screens, endosomal trafficking studies, and actin cytoskeleton research. Representative assays that can be performed with this model include Western blotting to confirm loss of CCDC93 protein expression, immunofluorescence to visualize endosomal tubulation defects, transferrin recycling assays coupled with flow cytometry to quantify cargo sorting efficiency, and co-immunoprecipitation to assess WASH complex integrity. The knockout cells also enable targeted investigation of ITGB1 and TFRC recycling dynamics. Researchers can leverage this system to explore how endosomal trafficking defects contribute to diseases such as neurodegenerative disorders. For technical inquiries or further details, please contact Ascent Research.
