The HDLBP Knockout HEK293T Polyclonal Cells constitute a polyclonal knockout population generated by CRISPR/Cas9-mediated disruption of the HDLBP gene in the HEK293T human cell line. This product offers a diverse pool of edited cells, facilitating robust loss-of-function analysis while mitigating clonal selection biases inherent to monoclonal lines. The polyclonal format preserves genetic variability, making it well-suited for studying gene function in a near-physiological epithelial context.
HEK293T cells are derived from human embryonic kidney and have been immortalized with sheared adenovirus 5 DNA. They stably express the SV40 large T antigen, which facilitates high-level protein expression and efficient transfection. These epithelial cells are a foundational tool in biomedical research, widely used for studying gene expression, protein interactions, and cellular signaling pathways. Their robust growth and genetic manipulability make them an ideal host for CRISPR-mediated knockout studies.
HDLBP encodes a scavenger receptor for HDL that mediates selective cholesterol ester uptake and also functions as an RNA-binding protein. Its transcription is controlled by sterol-responsive transcription factors SREBF1, SREBF2, and NR1H3 (LXR??) in response to cholesterol levels. At the protein level, HDLBP binds APOA1, HDL particles, RNA, and the translation initiation factor EIF4G, linking it to both cholesterol trafficking and mRNA translation and stability. This positions HDLBP at a critical node intersecting lipid metabolism and post-transcriptional regulation, with downstream effects on HDL uptake, cellular cholesterol levels, and RNA turnover.
In the HEK293T model, knockout of HDLBP allows dissection of its dual roles in cholesterol homeostasis and RNA biology. This system is pertinent for investigating dyslipidemia, atherosclerosis, and metabolic syndrome, where HDL metabolism is disrupted. Loss of HDLBP can alter cellular lipid profiles, HDL binding capacity, and the stability of target mRNAs, thereby revealing mechanistic links between lipid transport and gene expression. Moreover, the epithelial context provides a relevant background for studying HDLBP’s role in selective lipid uptake, which is central to reverse cholesterol transport and atherogenesis.
Researchers can utilize this polyclonal knockout population in functional assays such as cholesterol uptake and HDL binding assays to quantify lipid transport, Western blotting and RT-qPCR to confirm gene disruption and monitor downstream targets, and RNA immunoprecipitation to explore RNA interactions. Transcriptomic profiling by RNA-seq and lipidomic analyses further elucidate the impact of HDLBP loss on cellular metabolism. These applications are valuable for studies of atherosclerosis, dyslipidemia, and RNA-binding protein biology. For more information, please contact Ascent Research.
