AMPKβ1 CRISPR Activation Plasmid (h): sc-402952-ACT

PRKAB1 encodes the β1 regulatory subunit of AMP-activated protein kinase (AMPK), a central cellular energy sensor that coordinates metabolic homeostasis in response to nutrient and stress signals. AMPKβ1 supports assembly and stability of the AMPK heterotrimer and contributes to substrate targeting and subcellular localization, thereby influencing phosphorylation programs that control glucose uptake, fatty acid oxidation, mitochondrial biogenesis, autophagy, and mTORC1 signaling. Through these pathways, AMPKβ1 impacts cell growth decisions, redox balance, and adaptation to hypoxia and energetic stress. Dysregulated AMPK signaling has been associated with metabolic disorders, inflammation, and cancer-relevant phenotypes, making PRKAB1 a useful node for mechanistic studies of energy-dependent signaling networks.

AMPKβ1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PRKAB1 expression without altering the underlying DNA sequence.

AMPKβ1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PRKAB1 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 PRKAB1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous AMPKβ1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PRKAB1 locus and enabling the study of AMPKβ1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of AMPKβ1 pathway restoration in tumor cells with silenced or reduced PRKAB1 expression.

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