PGC1a CRISPR Activation Plasmid (m): sc-422364-ACT

Ppargc1a encodes the transcriptional coactivator PGC1a, a central regulator of mitochondrial biogenesis and oxidative metabolism in mouse tissues with high energetic demand. PGC1a coordinates nuclear receptor and transcription factor programs, including PPAR, NRF1/2, and ERR signaling, to tune fatty acid oxidation, oxidative phosphorylation, and antioxidant defenses. Its activity integrates nutrient and stress cues through pathways such as AMPK and sirtuin-dependent deacetylation, shaping metabolic flexibility and cellular redox balance. Dysregulated Ppargc1a expression has been linked to metabolic syndrome phenotypes, muscle and cardiac remodeling, and neurodegeneration-associated mitochondrial dysfunction, making it a frequent target in mechanistic studies of energy homeostasis.

PGC1a CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Ppargc1a expression without altering the underlying DNA sequence.

PGC1a CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Ppargc1a 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 Ppargc1a transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PGC1a expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Ppargc1a locus and enabling the study of PGC1a-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PGC1a pathway restoration in tumor cells with silenced or reduced Ppargc1a expression.

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