PIASx CRISPR Activation Plasmid (h): sc-403954-ACT

Human PIAS2 encodes PIASx, a PIAS family E3 SUMO ligase that modulates transcription factor activity and protein stability through SUMOylation. PIASx regulates signal transduction and gene expression programs linked to JAK/STAT signaling, nuclear receptor pathways, and DNA damage–responsive transcriptional control, thereby influencing cell cycle progression and stress adaptation. By tuning the output of cytokine and hormone-dependent transcriptional networks, PIASx contributes to context-specific regulation of proliferation and differentiation. Dysregulated SUMO pathway activity and altered PIAS2 expression have been associated with aberrant transcriptional states observed in cancer-related biology and inflammatory signaling models.

PIASx CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PIAS2 expression without altering the underlying DNA sequence.

PIASx CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PIAS2 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 PIAS2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PIASx expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PIAS2 locus and enabling the study of PIASx-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PIASx pathway restoration in tumor cells with silenced or reduced PIAS2 expression.

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