AKAP 2 CRISPR Activation Plasmid (m2): sc-419065-ACT-2

Mouse Akap2 encodes A-kinase anchoring protein 2 (AKAP2), a scaffold that spatially organizes protein kinase A (PKA) and associated signaling components to confer compartment-specific cAMP responses. By recruiting PKA to defined subcellular sites, AKAP2 influences phosphorylation networks that regulate cytoskeletal dynamics, cell polarity, membrane trafficking, and context-dependent transcriptional programs downstream of cAMP signaling. Disruption or altered regulation of AKAP-anchored PKA microdomains is relevant to studies of developmental signaling, neuronal and cardiomyocyte excitability, and broader mechanisms of signaling dysregulation implicated in disease-associated phenotypes. Akap2 gene-editing and functional genomics workflows enable interrogation of PKA compartmentalization, mapping of protein–protein interaction interfaces, and validation of pathway-dependent cellular readouts in mouse models and engineered cell systems.

AKAP 2 CRISPR Activation Plasmid (m2) provides a targeted, non-destructive approach to upregulating endogenous Akap2 expression without altering the underlying DNA sequence.

AKAP 2 CRISPR Activation Plasmid (m2) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Akap2 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 Akap2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous AKAP 2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Akap2 locus and enabling the study of AKAP 2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of AKAP 2 pathway restoration in tumor cells with silenced or reduced Akap2 expression.

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