PIASy CRISPR Activation Plasmid (h): sc-403125-ACT

Human PIAS4 encodes PIASy, a SUMO E3 ligase and transcriptional coregulator that modulates protein stability, subcellular localization, and transcription factor activity through SUMOylation. PIASy participates in stress and inflammatory signaling by regulating pathways including JAK/STAT and NF-κB, and it contributes to DNA damage responses by influencing the activity of key nuclear regulators. By shaping transcriptional programs linked to proliferation, apoptosis, and immune responses, PIASy has been implicated in contexts relevant to oncogenic signaling, immune dysregulation, and genome stability. Its central role in post-translational modification makes PIAS4 a useful node for mechanistic studies of SUMO-dependent control of nuclear signaling networks.

PIASy CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PIAS4 expression without altering the underlying DNA sequence.

PIASy CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PIAS4 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 PIAS4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PIASy expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PIAS4 locus and enabling the study of PIASy-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PIASy pathway restoration in tumor cells with silenced or reduced PIAS4 expression.

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