Ptx3 CRISPR Activation Plasmid (h): sc-402766-ACT

PTX3 encodes pentraxin 3 (Ptx3), a soluble pattern-recognition molecule of the long pentraxin family that functions in innate immunity and extracellular matrix regulation. Ptx3 is rapidly induced by inflammatory cues such as IL-1, TNF, and microbial products, and contributes to complement activation control, opsonization, and modulation of leukocyte recruitment within NF-κB– and cytokine-driven responses. It also participates in tissue remodeling by binding matrix components and growth factors, linking inflammatory signaling to angiogenesis and stromal organization. Dysregulated PTX3 expression is associated with chronic inflammatory states, infection biology, cardiovascular and metabolic inflammation, and tumor-associated microenvironment dynamics, supporting its use as a functional readout in immunology and disease-model studies.

Ptx3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PTX3 expression without altering the underlying DNA sequence.

Ptx3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PTX3 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 PTX3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Ptx3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PTX3 locus and enabling the study of Ptx3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Ptx3 pathway restoration in tumor cells with silenced or reduced PTX3 expression.

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