The CCDC138 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the near-haploid human HAP1 cell line, with targeted disruption of the CCDC138 gene. This loss-of-function model enables investigation of CCDC138, a coiled-coil protein critical for sperm flagellum assembly and ciliogenesis. The polyclonal format provides a heterogeneous pool of edited alleles for robust functional studies without clonal selection artifacts.
HAP1 cells originate from the KBM-7 chronic myeloid leukemia (CML) cell line, established from a male patient in blast crisis, and exhibit a near-haploid karyotype that simplifies genetic analyses. As immortalized leukemic myeloid progenitor cells, HAP1 cells retain key features of hematopoietic lineage while providing a stable, proliferative model for high-throughput perturbation screens. The near-haploid genome facilitates CRISPR-based knockout studies by reducing gene copy-number complications, making it a preferred platform for systematic investigation of gene function.
CCDC138 encodes a coiled-coil protein localizing to the manchette and sperm flagellum, essential for axoneme assembly and flagellar motility. It functions downstream of transcription factors RFX2 and FOXJ1, which regulate ciliary gene expression. CCDC138 interacts with IFT proteins and centrosomal coiled-coil proteins like CEP135 and CEP250, facilitating assembly of axonemal dynein complexes and sperm flagellar structural proteins including DNAH1 and AKAP4. Loss of CCDC138 disrupts these interactions, causing multiple morphological abnormalities of the sperm flagella (MMAF) and male infertility. CCDC138 also contributes to centrosome duplication and organization.
In HAP1 cells, the polyclonal CCDC138 knockout population serves as a powerful tool for dissecting centrosomal and ciliary functions. Despite its well-established role in spermatogenesis, the near-haploid background of HAP1 cells enables unambiguous assessment of CCDC138??s involvement in centrosome organization and ciliogenesis, processes often dysregulated in leukemia and other cancers. The simplified genetic landscape allows for precise mapping of genetic interactions and facilitates the identification of synthetic lethal relationships in drug discovery contexts.
This knockout model supports diverse assays: western blotting for CCDC138, immunofluorescence for centrosomal (??-tubulin) and ciliary (acetylated tubulin) markers, RT-qPCR for ciliogenic gene expression, and co-immunoprecipitation with IFT and centrosomal components. Applications include functional analyses of CCDC138 in centrosome and cilia biology, elucidation of RFX2/FOXJ1 signaling, high-throughput genetic screens for ciliary modifiers, and drug target discovery for male infertility. For technical support, contact Ascent Research.
