P2X2 CRISPR Activation Plasmid (h): sc-403517-ACT

Human P2RX2 encodes the ATP-gated P2X2 receptor, a trimeric nonselective cation channel that mediates rapid Na⁺/Ca²⁺ influx in response to extracellular nucleotides. P2X2 signaling shapes membrane excitability, Ca²⁺-dependent second messenger pathways, and activity-dependent transcription in neurons and sensory epithelia, influencing synaptic transmission, neurosecretion, and sensory processing. This purinergic receptor participates in ATP-driven communication during tissue stress and inflammation, integrating with broader neuroimmune signaling networks. Altered P2RX2/P2X2 function has been associated with sensory and neurologic phenotypes, supporting its study in mechanisms underlying auditory and nociceptive signaling and related channelopathy-relevant pathways.

P2X2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous P2RX2 expression without altering the underlying DNA sequence.

P2X2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the P2RX2 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 P2RX2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous P2X2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native P2RX2 locus and enabling the study of P2X2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of P2X2 pathway restoration in tumor cells with silenced or reduced P2RX2 expression.

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