The HDLBP Knockout HAP1 Polyclonal Cells are a ready-to-use CRISPR/Cas9-edited polyclonal knockout population in the near-haploid HAP1 cell line, generated by Cas9-mediated gene disruption of the HDLBP locus. This pool of edited cells provides a robust and versatile model for functional genomics studies, enabling loss-of-function analysis without the need for clonal isolation.
The HAP1 host cell line is a human near-haploid line derived from a chronic myeloid leukemia patient, exhibiting fibroblast-like morphology and a predominantly haploid karyotype. Its genetic simplicity allows efficient single-allele disruption, making it ideal for CRISPR-based genetic screens, drug target validation, and functional genomics.
HDLBP, also known as vigilin, is a conserved RNA-binding protein that directly binds high-density lipoprotein (HDL) particles and regulates the stability and translation of mRNAs encoding cholesterol efflux transporters. It specifically interacts with and stabilizes ABCA1 and ABCG1 mRNAs, promoting reverse cholesterol transport. Upstream, HDLBP expression is induced by LXR agonists and SREBP1, while downstream, it facilitates the lipidation of APOA1 to form mature HDL. Additionally, HDLBP can associate with AGO2 in the RNA-induced silencing complex, implicating it in microRNA-mediated regulation. These molecular interactions establish HDLBP as a central player in cholesterol homeostasis and mRNA metabolism.
In the HAP1 cellular context, disruption of HDLBP provides a powerful platform to dissect cholesterol trafficking and lipid metabolism. The near-haploid background minimizes genetic redundancy, ensuring clear phenotypic outcomes. This model is particularly relevant for studying metabolic disorders such as non-alcoholic fatty liver disease, cardiovascular disease, and metabolic syndrome, where HDLBP dysfunction may lead to pathogenic lipid accumulation. It also serves as a tool for drug target discovery in these conditions.
Researchers can employ these polyclonal knockout cells in a broad range of functional assays: Western blotting to confirm HDLBP depletion, RT-qPCR to quantify ABCA1 and ABCG1 mRNA levels, and cholesterol efflux assays to measure HDL-mediated lipid transport. Fluorescent HDL uptake assays and Oil Red O staining enable visualization of lipid accumulation. RNA immunoprecipitation (RIP) can assess HDLBP?CRNA interactions, while RNA-seq provides transcriptome-wide insights into mRNA metabolism. The polyclonal format is ideal for pooled CRISPR modifier screens. For additional information on ordering and technical support, please contact Ascent Research.
