PIAS 3 CRISPR Activation Plasmid (h): sc-401406-ACT

PIAS3 (protein inhibitor of activated STAT3) is a SUMO E3 ligase that modulates transcriptional responses by promoting SUMOylation of nuclear proteins and by directly inhibiting DNA-binding activity of STAT3. Through regulation of JAK/STAT signaling and broader SUMO-dependent control of transcription factor activity, PIAS3 influences cytokine signaling outputs, cell-cycle progression, and stress-responsive gene programs. PIAS3 has been linked to pathways governing immune regulation and oncogenic signaling, where altered STAT3 activity and SUMOylation dynamics are frequently observed. Studying PIAS3 function helps clarify how post-translational modification and transcriptional repression shape gene expression states in inflammation-associated and proliferative contexts.

PIAS 3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous expression without altering the underlying DNA sequence.

PIAS 3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the 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 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PIAS 3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native locus and enabling the study of PIAS 3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PIAS 3 pathway restoration in tumor cells with silenced or reduced expression.

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