AMPKγ2 CRISPR Activation Plasmid (h): sc-404970-ACT

PRKAG2 encodes the AMPKγ2 regulatory subunit of AMP-activated protein kinase (AMPK), a central energy sensor that couples cellular AMP/ATP status to metabolic adaptation. By modulating AMPK complex assembly and nucleotide sensing, AMPKγ2 influences downstream pathways controlling glucose uptake, fatty acid oxidation, mitochondrial biogenesis, and autophagy through signaling nodes such as mTORC1 and ULK1. PRKAG2 is highly relevant to cardiac and skeletal muscle bioenergetics, where altered AMPK signaling can shift substrate utilization and stress responses. Genetic variation or dysregulation of PRKAG2 has been associated with cardiac conduction and hypertrophic phenotypes, supporting its use in mechanistic studies of metabolic and electrophysiological remodeling.

AMPKγ2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PRKAG2 expression without altering the underlying DNA sequence.

AMPKγ2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PRKAG2 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 PRKAG2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous AMPKγ2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PRKAG2 locus and enabling the study of AMPKγ2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of AMPKγ2 pathway restoration in tumor cells with silenced or reduced PRKAG2 expression.

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