The KCTD20 Knockout HAP1 Polyclonal Cells product consists of a polyclonal knockout cell population generated by CRISPR/Cas9-mediated disruption of the KCTD20 gene in the HAP1 human near-haploid chronic myeloid leukemia (CML) cell line. This polyclonal pool comprises a heterogeneous mixture of gene-edited cells, each carrying distinct loss-of-function mutations in KCTD20, providing a robust model for studying gene function without clonal selection. The polyclonal format preserves genetic diversity and is suitable for pooled screening applications, offering a cost-effective and scalable tool for investigating the KCTD20-dependent ubiquitin-proteasome pathway in cancer cell biology.
The HAP1 host cell line is a widely adopted model system for functional genomics, derived from the KBM-7 CML patient cell line and possessing a near-haploid karyotype. This unique genetic background facilitates efficient gene targeting and phenotype analysis, as the presence of a single allele simplifies the establishment of knockout genotypes. HAP1 cells retain key features of the myeloid lineage and are widely used in CRISPR screens, protein interaction studies, and drug target validation. Their adherent growth and robust proliferation make them suitable for a variety of cell-based assays.
KCTD20 encodes a putative substrate recognition receptor for the Cullin3-RING E3 ubiquitin ligase complex, a central component of the ubiquitin-proteasome system. Within this complex, KCTD20 likely acts as an adaptor that recruits specific protein substrates to the Cullin3 scaffold, enabling their ubiquitination by the RBX1-associated E2 enzyme and subsequent degradation by the 26S proteasome. KCTD20 function is regulated by the NEDD8 conjugation pathway, which activates Cullin3 through neddylation, and its activity is mechanistically linked to protein quality control and cell cycle progression. While its downstream targets remain largely uncharacterized, KCTD20 is hypothesized to mediate turnover of key regulatory proteins.
Loss of KCTD20 in the HAP1 near-haploid background facilitates unambiguous assessment of its role in ubiquitin-dependent degradation pathways. This knockout model allows researchers to dissect the contribution of KCTD20 to Cullin3-RING ligase activity and to identify its endogenous substrates using comparative proteomics. The HAP1 system??s genetic simplicity reduces confounding effects from paralog compensation, making it particularly valuable for mapping functional domains and for screening chemical modulators of the ubiquitin-proteasome system.
Typical applications include genomic loss-of-function screens, co-immunoprecipitation studies to validate Cullin3 interaction, ubiquitination assays, quantitative proteomics for substrate identification, and flow cytometric analysis of cell cycle and apoptosis markers. Combined with Western blotting and RT-qPCR for confirmation, this polyclonal knockout pool supports detailed mechanistic studies into KCTD20 biology. For ordering information and technical support, please contact Ascent Research.
