PDE7B CRISPR Activation Plasmid (h): sc-411961-ACT

Human PDE7B encodes phosphodiesterase 7B, a cAMP-specific phosphodiesterase that hydrolyzes cyclic AMP to 5′-AMP and thereby shapes the amplitude and duration of intracellular cAMP signaling. By controlling cAMP-dependent pathways such as PKA/CREB-mediated transcription and downstream modulation of immune and neuronal signaling programs, PDE7B contributes to regulation of cellular activation, differentiation, and survival. PDE7B expression and activity have been linked to inflammatory signaling networks and neurobiology-relevant processes, making it a useful node for studying cAMP compartmentalization and signal integration. Dysregulation of PDE7B-associated signaling has been reported in contexts relevant to immune dysfunction and CNS-related phenotypes, supporting its investigation as a pathway modulator in disease models.

PDE7B CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PDE7B expression without altering the underlying DNA sequence.

PDE7B CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PDE7B 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 PDE7B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PDE7B expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PDE7B locus and enabling the study of PDE7B-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PDE7B pathway restoration in tumor cells with silenced or reduced PDE7B expression.

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