AMPK alpha 1 CRISPR Activation Plasmid (m2): sc-430618-ACT-2

Mouse Prkaa1 encodes the catalytic α1 subunit of AMP-activated protein kinase (AMPKα1), a central energy sensor that is activated by rising AMP/ADP levels to restore ATP homeostasis. AMPKα1 integrates upstream inputs from LKB1 and CaMKK2 and regulates metabolic pathways including fatty acid oxidation, glucose uptake, and inhibition of anabolic programs via targets such as ACC and mTORC1, while also influencing autophagy and mitochondrial biogenesis. Dysregulation of AMPKα1 signaling is linked to altered nutrient sensing and cellular stress responses relevant to obesity, type 2 diabetes, hepatic steatosis, inflammation, and tumor metabolism. Prkaa1 gene editing in mouse models enables mechanistic studies of energy balance, immune cell activation and polarization, and tissue-specific metabolic remodeling, supporting functional genomics workflows in metabolic and cancer biology research.

AMPK alpha 1 CRISPR Activation Plasmid (m2) provides a targeted, non-destructive approach to upregulating endogenous Prkaa1 expression without altering the underlying DNA sequence.

AMPK alpha 1 CRISPR Activation Plasmid (m2) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Prkaa1 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 Prkaa1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous AMPK alpha 1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Prkaa1 locus and enabling the study of AMPK alpha 1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of AMPK alpha 1 pathway restoration in tumor cells with silenced or reduced Prkaa1 expression.

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