PDE2A CRISPR Activation Plasmid (h): sc-401957-ACT

Human PDE2A encodes phosphodiesterase 2A, a dual-substrate cyclic nucleotide phosphodiesterase that hydrolyzes cAMP and cGMP and couples nitric oxide–cGMP signals to cAMP-dependent pathways. By shaping cyclic nucleotide gradients, PDE2A modulates PKA/PKG signaling, calcium handling, and downstream transcriptional programs that influence neuronal excitability and synaptic plasticity, as well as vascular and cardiac responses. PDE2A activity is implicated in regulation of mitochondrial and inflammatory stress responses through cAMP/CREB and related signaling nodes. Dysregulated cyclic nucleotide turnover involving PDE2A has been associated with neuropsychiatric and neurodegenerative phenotypes and cardiometabolic dysfunction, supporting its use as a target for pathway dissection in disease-relevant models.

PDE2A CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PDE2A expression without altering the underlying DNA sequence.

PDE2A CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PDE2A 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 PDE2A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PDE2A expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PDE2A locus and enabling the study of PDE2A-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PDE2A pathway restoration in tumor cells with silenced or reduced PDE2A expression.

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