P2Y2 CRISPR Activation Plasmid (m): sc-422096-ACT

Mouse P2ry2 encodes the purinergic receptor P2Y2 (P2Y2R), a G protein–coupled receptor activated by extracellular ATP and UTP that couples primarily to Gq/11 to stimulate phospholipase C, intracellular Ca2+ mobilization, and PKC-dependent signaling. P2Y2 also engages MAPK/ERK and PI3K-related pathways and can modulate cytoskeletal remodeling and integrin-linked migration, supporting roles in epithelial transport, mechanosensory responses, and leukocyte trafficking. In immune and barrier tissues, P2Y2 signaling contributes to chemokine production and inflammasome-adjacent stress responses driven by nucleotides released during tissue injury. Dysregulated purinergic signaling involving P2Y2 has been studied in inflammatory lung disease, chronic airway remodeling, pain and neuroinflammation, and tumor-associated microenvironmental signaling, making P2ry2 a useful target for pathway and phenotypic interrogation in mouse models.

P2Y2 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous P2ry2 expression without altering the underlying DNA sequence.

P2Y2 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the P2ry2 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 P2ry2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous P2Y2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native P2ry2 locus and enabling the study of P2Y2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of P2Y2 pathway restoration in tumor cells with silenced or reduced P2ry2 expression.

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