LDLR CRISPR Activation Plasmid (h): sc-400645-ACT

Human LDLR encodes the low-density lipoprotein receptor (LDLR), a cell-surface endocytic receptor that binds ApoB- and ApoE-containing lipoproteins to mediate clathrin-dependent internalization and lysosomal processing of cholesterol-rich particles. LDLR activity is a central determinant of cellular cholesterol homeostasis and feeds into SREBP-regulated lipid biosynthesis and uptake programs, integrating with sterol sensing, membrane trafficking, and endosome–lysosome pathways. Perturbation of LDLR expression or function is strongly linked to dyslipidemia biology and atherosclerosis-relevant mechanisms, making it widely used to study lipoprotein metabolism, receptor recycling, and lipid-driven signaling responses. LDLR is also leveraged as a readout gene for transcriptional control of cholesterol pathways and for modeling genotype–phenotype relationships in lipid handling.

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

LDLR CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the LDLR 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 LDLR transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LDLR expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native LDLR locus and enabling the study of LDLR-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LDLR pathway restoration in tumor cells with silenced or reduced LDLR expression.

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