GPIHBP1 CRISPR Activation Plasmid (h): sc-403473-ACT

GPIHBP1 (glycosylphosphatidylinositol anchored high density lipoprotein binding protein 1) is an endothelial cell surface GPI-anchored protein that captures lipoprotein lipase (LPL) and transports it to the capillary lumen, enabling intravascular hydrolysis of triglyceride-rich lipoproteins. Through its binding to LPL and interactions with chylomicrons and VLDL, GPIHBP1 is central to the lipolysis pathway and the regulation of plasma triglyceride clearance. Altered GPIHBP1 expression or function perturbs LPL localization and activity, contributing to dyslipidemia phenotypes characterized by hypertriglyceridemia and lipid handling defects. This gene is therefore widely studied in vascular lipid metabolism, endothelial biology, and mechanisms governing lipoprotein processing.

GPIHBP1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GPIHBP1 expression without altering the underlying DNA sequence.

GPIHBP1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GPIHBP1 locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the GPIHBP1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GPIHBP1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GPIHBP1 locus and enabling the study of GPIHBP1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GPIHBP1 pathway restoration in tumor cells with silenced or reduced GPIHBP1 expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.