CPTI CRISPR Activation Plasmid (m): sc-419786-ACT

Mouse Cpt1a encodes carnitine palmitoyltransferase 1A (CPTI), the rate-limiting enzyme that controls mitochondrial import of long-chain fatty acids for β-oxidation via the carnitine shuttle. By regulating acetyl-CoA production and energy flux, CPTI integrates nutrient availability with mitochondrial respiration and influences lipid homeostasis, redox balance, and metabolic signaling. Cpt1a activity shapes cellular responses to fasting and high-fat conditions and contributes to metabolic remodeling in tissues with high oxidative demand. Dysregulation of CPTI-linked fatty acid oxidation has been associated with metabolic phenotypes relevant to insulin sensitivity, hepatic lipid handling, and inflammatory activation states in mouse models.

CPTI CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Cpt1a expression without altering the underlying DNA sequence.

CPTI CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Cpt1a 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 Cpt1a transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CPTI expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Cpt1a locus and enabling the study of CPTI-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CPTI pathway restoration in tumor cells with silenced or reduced Cpt1a expression.

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