The BTN1A1 Knockout HAP1 Polyclonal Cells product comprises a heterogeneous population of HAP1 cells with CRISPR/Cas9-mediated disruption of the BTN1A1 gene. As a polyclonal knockout pool, this product provides a versatile loss-of-function model for studying BTN1A1-dependent processes, free from the clonal biases of single-cell-derived lines.
HAP1 is a human near-haploid chronic myeloid leukemia-derived cell line, originally derived from KBM-7 and adapted for adherent growth. Its near-haploid karyotype facilitates efficient gene disruption, making it ideal for functional genomics and genetic screens. In a haploid background, CRISPR/Cas9-mediated gene editing yields unambiguous knockouts, enabling clear phenotypic analyses without interference from a second allele.
BTN1A1, a butyrophilin family member, is essential for milk fat globule secretion in mammary epithelial cells by interacting with xanthine oxidoreductase (XDH) at the apical membrane. This interaction is crucial for the envelopment and release of lipid droplets. BTN1A1 expression is regulated by prolactin and glucocorticoids via the prolactin receptor?CJAK2?CSTAT5 signaling axis, with STAT5 directly activating transcription. Beyond lactation, BTN1A1 may modulate immune responses through its B7-like extracellular domains, although its immunological roles are not fully characterized.
In HAP1 cells, the BTN1A1 knockout model offers a simplified system to dissect butyrophilin-mediated signaling and protein interactions. Although HAP1 cells do not naturally secrete milk fat, they can be used to study conserved pathways such as STAT5 activation and XDH binding, or they can be engineered to express mammary factors. The haploid background ensures that observed phenotypes are directly attributable to BTN1A1 loss, enhancing the utility of this model in high-throughput screens for modulators of butyrophilin function or lactation-related pathways.
This product is suited for functional studies of milk fat secretion, immune modulation by butyrophilins, and genome-wide screens for lactation phenotypes. Validation assays include Sanger sequencing for indel confirmation, western blotting for protein knockout, RT-qPCR for mRNA expression, and co-immunoprecipitation to assess XDH interaction. Immunofluorescence can reveal localization changes, while engineered mammary cell models enable milk fat globule secretion assays. For additional details, please contact Ascent Research.
