PKA IIα reg CRISPR Activation Plasmid (h): sc-401080-ACT

PRKAR2A encodes the regulatory subunit IIα of cAMP-dependent protein kinase A (PKA), a key modulator of kinase activity that constrains catalytic subunits in the absence of cAMP and tunes signaling amplitude and compartmentalization. Through anchoring interactions with AKAP scaffolds, PRKAR2A helps localize PKA to defined subcellular microdomains, shaping phosphorylation programs that control metabolism, cell-cycle progression, transcriptional responses, and synaptic signaling. This cAMP/PKA axis intersects with GPCR signaling and CREB-dependent gene regulation, influencing differentiation and stress-response pathways. Dysregulated PKA signaling and altered regulatory subunit balance have been associated with aberrant growth control and pathway rewiring observed in multiple disease contexts, supporting mechanistic studies of signal transduction and cellular plasticity.

PKA IIα reg CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PRKAR2A expression without altering the underlying DNA sequence.

PKA IIα reg CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PRKAR2A 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 PRKAR2A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PKA IIα reg expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PRKAR2A locus and enabling the study of PKA IIα reg-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PKA IIα reg pathway restoration in tumor cells with silenced or reduced PRKAR2A expression.

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