UCP3 CRISPR Activation Plasmid (h): sc-400866-ACT

Human UCP3 (uncoupling protein 3) is a mitochondrial inner membrane carrier that modulates proton leak and influences coupling efficiency of oxidative phosphorylation, shaping ATP production, reactive oxygen species balance, and heat generation in metabolically active tissues. It is strongly linked to fatty acid oxidation and mitochondrial quality control, with roles in regulating substrate utilization during nutrient stress and high lipid flux. Altered UCP3 expression has been associated with insulin resistance, obesity-linked metabolic dysfunction, and skeletal muscle mitochondrial phenotypes, supporting its use as a node for studying energy homeostasis. UCP3 is therefore relevant to pathways integrating mitochondrial respiration, lipid handling, and redox signaling in cellular and organismal metabolism.

UCP3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous UCP3 expression without altering the underlying DNA sequence.

UCP3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the UCP3 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 UCP3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous UCP3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native UCP3 locus and enabling the study of UCP3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of UCP3 pathway restoration in tumor cells with silenced or reduced UCP3 expression.

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