The CATSPER2 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the near-haploid HAP1 cell line. This loss-of-function model disrupts CATSPER2, which encodes a pore-forming subunit of the sperm-specific CatSper calcium channel. The polyclonal format captures diverse loss-of-function alleles, avoiding clonal selection bias and providing a robust tool for high-throughput screening applications in a genetically tractable background.
HAP1 is a near-haploid subclone of the CML-derived KBM-7 cell line, favored for functional genomics because its haploid genome minimizes genetic redundancy and enables unambiguous genotype?Cphenotype linkages. It retains key signaling pathways, including calcium and kinase cascades, and offers high transfection efficiency and rapid proliferation, facilitating efficient CRISPR editing and multiplexed phenotypic analyses.
CATSPER2 encodes a pore-forming ?? subunit of the CatSper complex, a voltage-gated calcium channel that mediates sperm hyperactivation. Channel opening is triggered by progesterone or intracellular alkalinization, leading to Ca2+ influx and activation of calmodulin (CaM), Ca2+/calmodulin-dependent kinase II (CaMKII), calcineurin, and PKA. CATSPER2 interacts with auxiliary subunits CATSPER1, CATSPER3, CATSPER4, CATSPER??, and CATSPER??, as well as with ANO1 and SLC9B2. These interactions are critical for channel assembly and function.
In the HAP1 model, CATSPER2 knockout enables dissection of channel assembly and signaling without germline-specific constraints. The near-haploid genome simplifies protein?Cprotein interaction studies, allowing co-immunoprecipitation and functional complementation assays. This system is particularly suited for calcium imaging and high-content screening, facilitating validation of infertility-associated mutations and identification of channel modulators.
Applications include CRISPR-based screening for CatSper regulators, functional validation of clinical CATSPER2 variants, and drug target assessment for male fertility. Confirmatory assays include Western blotting, RT-qPCR, and Sanger indel analysis; functional studies use calcium imaging and immunofluorescence; protein interactions can be probed by co-immunoprecipitation and flow cytometry. For further information or to discuss your research needs, contact Ascent Research.
