The CATSPER1 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited human cell population with targeted disruption of the CATSPER1 gene in the near-haploid HAP1 cell line. Comprising a heterogeneous pool of loss-of-function alleles, these polyclonal knockout cells provide a renewable resource for investigating the absence of CATSPER1 protein via western blotting and for confirming genomic edits through PCR and Sanger sequencing. The product is delivered as a live culture suitable for immediate expansion in suspension, facilitating integration into workflows requiring calcium channel studies or drug screening.
HAP1 is a near-haploid cell line derived from KBM-7 chronic myeloid leukemia cells, adapted to suspension growth and extensively used for genetic knockout screening owing to its single-copy genome. The stable haploid karyotype (aside from a disomic chromosome 8) enables unambiguous disruption of target genes, making HAP1 an efficient platform for generating polyclonal knockout models. Its myeloid leukemia origin confers rapid proliferation and compatibility with high-throughput assays such as viability screens and drug sensitivity testing.
CATSPER1 encodes the pore-forming ?? subunit of the sperm-specific CATSPER calcium channel, activated by progesterone via SRC kinases, intracellular alkalinization, and cAMP-PKA signaling, with testosterone modulating its expression. The channel assembles with auxiliary subunits (CATSPER2, 3, 4, ??, ??, ??, ??) and associated proteins EFCAB9 and C2CD6 to mediate calcium influx into sperm flagella. Downstream, elevated calcium activates calmodulin and CaMKIV, driving hyperactivated motility and acrosome reaction, essential for male fertility.
Since CATSPER1 expression is normally restricted to testis, the HAP1 knockout provides a clean human genetic background for studying channel function without endogenous interference. These cells are ideal for reconstitution experiments, allowing co-expression of CATSPER subunits to investigate assembly and trafficking, and for antibody validation as a negative control. The haploid state simplifies interpretation of results, as re-introduction of wild-type CATSPER1 can assess functional rescue, while the knockout pool enables pooled screening assays.
Applications include male contraceptive drug screening by testing CATSPER channel blockers in a human cell context; CRISPR-editing validation through sequencing of the polyclonal pool; and studies of calcium signaling machinery or off-target effects of channel modulators. Standard assays such as RT-qPCR, immunofluorescence, calcium imaging (with channel reconstitution), and patch-clamp electrophysiology are readily applicable. For more information, please contact Ascent Research.
