PKA IIβ reg CRISPR Activation Plasmid (h): sc-402536-ACT

PRKAR2B encodes the regulatory subunit IIβ of cAMP-dependent protein kinase A (PKA), a key modulator of catalytic subunit activity and subcellular localization in response to cAMP. Through compartmentalized signaling often coordinated by A-kinase anchoring proteins (AKAPs), PRKAR2B helps shape phosphorylation dynamics that influence metabolism, cytoskeletal organization, transcriptional programs, and synaptic signaling. PKA-dependent pathways intersect with GPCR signaling, CREB-mediated gene regulation, and stress-responsive networks that control cellular proliferation and differentiation. Dysregulated cAMP/PKA signaling involving PRKAR2B has been linked in the literature to altered metabolic phenotypes and neurobehavioral processes, supporting its relevance for studying signal integration in disease-associated contexts.

PKA IIβ reg CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PRKAR2B expression without altering the underlying DNA sequence.

PKA IIβ reg CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PRKAR2B 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 PRKAR2B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PKA IIβ reg expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PRKAR2B locus and enabling the study of PKA IIβ reg-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PKA IIβ reg pathway restoration in tumor cells with silenced or reduced PRKAR2B expression.

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