UCP1 CRISPR Activation Plasmid (h): sc-400410-ACT

UCP1 (uncoupling protein 1) is a mitochondrial inner membrane proton transporter that dissipates the proton motive force to uncouple oxidative phosphorylation from ATP synthesis, increasing energy expenditure as heat. In humans, UCP1 is a defining component of brown and beige adipocyte thermogenic programs and influences mitochondrial respiration, fatty acid oxidation, and reactive oxygen species handling. Its transcriptional control integrates adrenergic signaling, PPAR/PGC-1α coactivator pathways, and broader metabolic stress responses that regulate adipose tissue remodeling. Altered UCP1 expression and activity are frequently studied in the context of obesity, insulin resistance, and cardiometabolic phenotypes, where thermogenic capacity and mitochondrial function are key variables.

UCP1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous UCP1 expression without altering the underlying DNA sequence.

UCP1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the UCP1 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 UCP1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous UCP1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native UCP1 locus and enabling the study of UCP1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of UCP1 pathway restoration in tumor cells with silenced or reduced UCP1 expression.

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