Cox-2 CRISPR Activation Plasmid (h): sc-400072-ACT

PTGS2 encodes cyclooxygenase-2 (Cox-2), an inducible, endoplasmic reticulum– and nuclear envelope–associated enzyme that converts arachidonic acid into prostaglandin H2, the committed precursor for diverse prostanoids. Cox-2 is rapidly upregulated by inflammatory cytokines, growth factors, and oncogenic stimuli and integrates signaling from NF-κB, MAPK/ERK, JAK/STAT, and AP-1 to modulate inflammatory gene programs. Through prostaglandin E2 and related lipid mediators, PTGS2 influences vascular tone, platelet function, pain and fever responses, epithelial barrier remodeling, and immune cell recruitment. Dysregulated PTGS2 expression is frequently observed in chronic inflammation and multiple cancer contexts, where it is linked to altered cell proliferation, angiogenic signaling, and immune evasion mechanisms.

Cox-2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PTGS2 expression without altering the underlying DNA sequence.

Cox-2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PTGS2 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 PTGS2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Cox-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PTGS2 locus and enabling the study of Cox-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Cox-2 pathway restoration in tumor cells with silenced or reduced PTGS2 expression.

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