CATSPER3 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the near-haploid HAP1 human cell line, designed for functional studies of CATSPER3. This gene encodes a pore-forming subunit of the sperm-specific CatSper calcium channel essential for hyperactivated sperm motility and male fertility. The product provides a heterogeneous pool of cells with CRISPR/Cas9-mediated disruption of the target gene, enabling loss-of-function analyses in a genetically tractable near-haploid background.
The HAP1 cell line originates from KBM-7 chronic myeloid leukemia cells and carries the BCR-ABL1 oncogenic fusion. Its near-haploid karyotype simplifies genetic engineering and makes it a widely adopted model for genetic screening and functional genomics. HAP1 cells grow adherently and maintain key signaling networks, offering a robust platform for probing gene function in cellular processes relevant to cancer biology and signal transduction.
CATSPER3 is an essential pore-forming subunit of the CatSper calcium channel, a sperm-specific complex that mediates calcium influx upon stimulation by upstream regulators such as progesterone, prostaglandin E1, and intracellular alkalinization. It assembles with CATSPER1, CATSPER2, CATSPER4, CATSPERB, and CATSPERG to form a functional channel. Downstream, this calcium entry triggers hyperactivated motility and acrosome reaction, processes critical for fertilization. Consequently, CATSPER3 disruption is linked to male infertility and oligoteratozoospermia, highlighting its role in calcium signaling pathways governing sperm function.
Although CATSPER3 expression is restricted to sperm, the HAP1 knockout model offers a valuable platform for studying CatSper channel assembly, calcium signaling mechanisms, and pharmacological modulation in a manipulable human cell system. Disruption of CATSPER3 eliminates the potential to form functional CatSper channels, enabling studies that reconstitute or compare channel composition and regulation. The model facilitates detailed examination of how progesterone, prostaglandin E1, and alkalinization influence channel activity and downstream calcium dynamics, contributing to the understanding of male infertility and ion channel-related pathologies.
These polyclonal knockout cells can be used in diverse assays to dissect CATSPER3 function and CatSper channel biology. Western blotting and RT-qPCR confirm loss of CATSPER3 expression, while immunofluorescence assesses protein localization in cells expressing channel components. RNA-seq enables transcriptome-wide profiling to identify changes associated with CatSper disruption. Functional calcium imaging following stimulation with progesterone or prostaglandin E1 measures channel activity. These applications support research in calcium signaling, male infertility, and ion channel-targeted therapeutics. For additional information, contact Ascent Research.
