PPARβ CRISPR Activation Plasmid (h): sc-400523-ACT

PPARD encodes peroxisome proliferator-activated receptor beta (PPARβ), a ligand-activated nuclear receptor that functions as a transcription factor regulating lipid handling, glucose homeostasis, and energy expenditure. PPARβ coordinates gene programs involved in fatty acid oxidation, mitochondrial function, and adaptive metabolic remodeling, integrating signals from lipid mediators to control cellular differentiation and inflammatory tone. In human tissues, PPARD activity is linked to skeletal muscle metabolism, keratinocyte biology, and macrophage polarization, intersecting with pathways such as PPAR signaling and broader nuclear receptor networks. Dysregulated PPARβ signaling has been associated with metabolic and inflammatory phenotypes and is frequently studied in contexts including insulin sensitivity, dyslipidemia, and tumor cell metabolic adaptation.

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

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

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