PIAS 1 CRISPR Activation Plasmid (h): sc-402407-ACT

PIAS1 encodes protein inhibitor of activated STAT1 (PIAS1), a SUMO E3 ligase and transcriptional coregulator that modulates signal-dependent gene expression. PIAS1 constrains cytokine and interferon responses by regulating STAT family activity and also influences NF-κB and p53-associated transcriptional programs through SUMOylation-dependent mechanisms. Through these functions it contributes to control of inflammation, DNA damage responses, and cell-cycle–linked transcriptional decisions. Dysregulated PIAS1 expression or activity has been associated with aberrant immune signaling and oncogenic transcriptional states, making it a useful node for pathway dissection in human cell models.

PIAS 1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PIAS1 expression without altering the underlying DNA sequence.

PIAS 1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PIAS1 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 PIAS1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PIAS 1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PIAS1 locus and enabling the study of PIAS 1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PIAS 1 pathway restoration in tumor cells with silenced or reduced PIAS1 expression.

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