PPARα CRISPR Activation Plasmid (h): sc-400238-ACT

PPARα (peroxisome proliferator-activated receptor alpha) is a ligand-activated nuclear receptor transcription factor that coordinates lipid catabolism and energy homeostasis in human cells. It regulates gene programs controlling mitochondrial and peroxisomal β-oxidation, fatty acid transport, ketogenesis, and lipoprotein metabolism, integrating metabolic cues with transcriptional outputs via PPAR response elements. PPARα activity intersects with AMPK and insulin signaling and modulates inflammatory gene expression through crosstalk with NF-κB and other nuclear receptors. Dysregulation of PPARA-associated pathways is frequently investigated in contexts such as metabolic syndrome, dyslipidemia, nonalcoholic fatty liver disease, and cardiometabolic remodeling, making it a central node for mechanistic studies of lipid-driven cellular stress.

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

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

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