AMPKβ2 CRISPR Activation Plasmid (h): sc-403537-ACT

PRKAB2 encodes the human AMPKβ2 regulatory subunit of AMP-activated protein kinase (AMPK), a central energy sensor that integrates nutrient availability with cellular metabolism. AMPKβ2 supports assembly and stability of the heterotrimeric AMPK complex and contributes to glycogen binding, enabling coordinated control of glucose uptake, glycolysis, fatty acid oxidation, and mitochondrial biogenesis. Through phosphorylation-dependent signaling, AMPK intersects with mTOR, autophagy, and insulin/IGF pathways to regulate growth, stress adaptation, and bioenergetic homeostasis. Dysregulated AMPK signaling and altered PRKAB2 expression have been associated with metabolic imbalance and cellular stress programs relevant to obesity, type 2 diabetes, and cancer metabolism research.

AMPKβ2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PRKAB2 expression without altering the underlying DNA sequence.

AMPKβ2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PRKAB2 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 PRKAB2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous AMPKβ2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PRKAB2 locus and enabling the study of AMPKβ2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of AMPKβ2 pathway restoration in tumor cells with silenced or reduced PRKAB2 expression.

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