AMPKγ1 CRISPR Activation Plasmid (h): sc-418058-ACT

PRKAG1 encodes the γ1 regulatory subunit of AMP-activated protein kinase (AMPK), a central cellular energy sensor that couples changes in AMP/ADP-to-ATP ratios to metabolic rewiring. AMPKγ1 contributes to nucleotide binding and allosteric regulation of the AMPK complex, influencing phosphorylation-driven control of glucose uptake, fatty acid oxidation, mitochondrial biogenesis, and autophagy. Through integration with mTORC1, insulin signaling, and stress-response pathways, AMPK activity helps coordinate cell growth with nutrient availability. Dysregulated AMPK signaling and altered PRKAG1 expression have been linked to metabolic dysfunction and may modulate tumor cell adaptation to energetic stress, making PRKAG1 a useful target for mechanistic studies of bioenergetics.

AMPKγ1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PRKAG1 expression without altering the underlying DNA sequence.

AMPKγ1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PRKAG1 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 PRKAG1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous AMPKγ1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PRKAG1 locus and enabling the study of AMPKγ1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of AMPKγ1 pathway restoration in tumor cells with silenced or reduced PRKAG1 expression.

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