PDE4D CRISPR Activation Plasmid (m): sc-433646-ACT

Mouse Pde4d encodes phosphodiesterase 4D (PDE4D), a cAMP-specific phosphodiesterase that hydrolyzes cAMP to 5′-AMP and shapes the amplitude and spatial confinement of PKA- and EPAC-dependent signaling. By regulating intracellular second-messenger tone, PDE4D influences GPCR-driven responses, transcriptional programs such as CREB signaling, and processes including cell proliferation, differentiation, and immune modulation. PDE4D activity is integrated with compartmentalized signaling via A-kinase anchoring proteins and can affect MAPK/ERK pathway dynamics through crosstalk with cAMP effectors. Dysregulated PDE4D-mediated cAMP turnover has been implicated in inflammation and neurobiological phenotypes, supporting its use as a mechanistic node in disease-relevant signaling studies in mouse models.

PDE4D CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Pde4d expression without altering the underlying DNA sequence.

PDE4D CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Pde4d 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 Pde4d transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PDE4D expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Pde4d locus and enabling the study of PDE4D-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PDE4D pathway restoration in tumor cells with silenced or reduced Pde4d expression.

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