PDE6β CRISPR Activation Plasmid (m): sc-422166-ACT

Mouse Pde6b encodes the beta subunit of cGMP phosphodiesterase 6 (PDE6β), a core effector of the rod phototransduction cascade. In retinal rod photoreceptors, PDE6β hydrolyzes cGMP downstream of light-activated rhodopsin–transducin signaling, thereby regulating cyclic nucleotide–gated channel activity, membrane hyperpolarization, and calcium-dependent adaptation processes. Proper PDE6β function is essential for maintaining photoreceptor homeostasis and signal fidelity, linking this enzyme to studies of sensory transduction, cGMP metabolism, and outer segment biology. Dysregulation or loss of PDE6β activity is widely used in experimental models of inherited retinal degeneration, enabling mechanistic analyses of photoreceptor stress responses, synaptic remodeling, and vision-related phenotypes.

PDE6β CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Pde6b expression without altering the underlying DNA sequence.

PDE6β CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Pde6b 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 Pde6b transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PDE6β expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Pde6b locus and enabling the study of PDE6β-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PDE6β pathway restoration in tumor cells with silenced or reduced Pde6b expression.

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