The CATSPER4 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population engineered to disrupt the human CATSPER4 gene within the near-haploid HAP1 fibroblast-like cell line. This polyclonal pool harbors a diverse array of loss-of-function mutations, enabling comprehensive gene inactivation and providing a versatile model for investigating CATSPER4-dependent channel biology, signaling pathways, and drug?Ctarget interactions. Researchers can leverage these cells for immunoblotting, immunofluorescence, and functional assays to validate target engagement and downstream effects.
HAP1 cells originate from the KBM-7 chronic myeloid leukemia line and possess a predominantly haploid genome, which facilitates CRISPR/Cas9-mediated gene disruption by eliminating the need for biallelic editing. This haploid background ensures complete functional knockout in the polyclonal population, as only a single targeted allele is present. The cells exhibit fibroblast-like morphology, vigorous growth, and broad compatibility with high-throughput screening platforms, making them an optimal host for genetic perturbation studies in cancer biology, signal transduction, and functional genomics.
CATSPER4 encodes a critical pore-forming subunit of the sperm-specific CatSper calcium channel, which is essential for hyperactivated motility and male fertility. The channel complex comprises additional pore-forming subunits (CatSper1?C3) and auxiliary subunits (CatSper??, ??, ??). Upstream activators such as progesterone, prostaglandin E1, and alkaline intracellular pH trigger CatSper opening via cAMP?CPKA signaling, leading to calcium influx. This calcium elevation promotes dynein ATPase activity and flagellar bending, driving the acrosome reaction and fertilization. Knockout of CATSPER4 abrogates channel assembly, eliminates calcium transients, and phenocopies CatSper-related male infertility.
In the HAP1 host, CATSPER4 knockout provides a null background for ectopic expression and reconstitution of the CatSper complex, allowing detailed structure?Cfunction analyses of subunit interactions, PKA-mediated phosphorylation, and channel trafficking. The polyclonal nature of the knockout population supports pooled screening applications and assessment of editing efficiency. Despite the lack of endogenous CATSPER4 expression in leukemic cells, these cells serve as a robust mammalian platform for co-immunoprecipitation, localization studies with fluorescently tagged subunits, and calcium imaging upon heterologous channel expression.
These polyclonal knockout cells are ideally suited for PCR genotyping and Western blot-based confirmation of CATSPER4 disruption, as well as for immunofluorescence localization of channel subunits. They provide a platform for calcium imaging experiments upon ectopic expression of CatSper channels, enabling real-time monitoring of calcium transients. The haploid background facilitates high-throughput drug screening for compounds that modulate channel assembly or function, supporting male contraceptive development. Co-immunoprecipitation and in vitro reconstitution assays benefit from the null background to map protein?Cprotein interactions within the CatSper complex without endogenous CATSPER4 interference. For additional product details or inquiries, contact Ascent Research.
