VLDLR CRISPR Activation Plasmid (h): sc-401310-ACT

VLDLR (very low density lipoprotein receptor) is a member of the LDL receptor family that binds apolipoprotein E–containing lipoproteins and the extracellular matrix glycoprotein reelin, linking endocytosis with signal transduction. In the reelin pathway, VLDLR functions with DAB1 phosphorylation to regulate neuronal migration, synaptic organization, and cortical layer formation. Outside the nervous system, VLDLR participates in lipid uptake and intracellular trafficking, influencing cellular metabolism and membrane homeostasis. Dysregulated VLDLR expression or signaling has been associated with neurodevelopmental phenotypes and altered lipid-handling states relevant to cardiometabolic and neurological research contexts.

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

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

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