apolipoprotein E/apoE CRISPR Activation Plasmid (h): sc-400860-ACT

APOE encodes apolipoprotein E (apoE), a secreted lipid-binding protein that mediates cholesterol and triglyceride transport through interactions with LDL receptor family members and heparan sulfate proteoglycans. ApoE is central to lipoprotein particle assembly and clearance and influences cellular lipid uptake, redistribution, and homeostatic signaling in hepatocytes, macrophages, and glial cells. In the nervous system, apoE modulates synaptic maintenance, neuroinflammatory responses, and proteostasis pathways that impact amyloid-β handling. Genetic and expression differences in APOE are strongly associated with dyslipidemia and neurodegeneration, making it a key target for mechanistic studies of lipid metabolism and brain immune–lipid crosstalk.

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

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

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