Pannexin-1 CRISPR Activation Plasmid (m): sc-424967-ACT

Mouse Panx1 encodes pannexin-1, a large-pore plasma membrane channel that mediates ATP release and purinergic signaling to coordinate intercellular communication. Pannexin-1 activity influences Ca2+ flux, inflammasome-related responses, and modulation of innate immune signaling, often intersecting with P2X/P2Y receptor pathways and pannexin channel gating mechanisms. In diverse cell types, PANX1 contributes to regulation of apoptosis-associated “find-me” signals, mechanosensitive responses, and inflammatory mediator release. Dysregulated PANX1 signaling has been linked in the literature to neuroinflammation, ischemic injury responses, and altered tumor–immune microenvironment signaling, supporting its study in disease-relevant models without implying clinical utility.

Pannexin-1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Panx1 expression without altering the underlying DNA sequence.

Pannexin-1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Panx1 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 Panx1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Pannexin-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Panx1 locus and enabling the study of Pannexin-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Pannexin-1 pathway restoration in tumor cells with silenced or reduced Panx1 expression.

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