Stat1 CRISPR Activation Plasmid (m): sc-423174-ACT

Signal transducer and activator of transcription 1 (Stat1) is a central transcription factor in mouse innate and adaptive immunity that transduces signals downstream of type I, II, and III interferons. Upon cytokine-induced JAK-mediated phosphorylation, STAT1 forms homo- or heterodimers, translocates to the nucleus, and binds GAS/ISRE elements to coordinate interferon-stimulated gene expression programs. These pathways regulate antiviral restriction, antigen presentation, macrophage polarization, and cell-cycle control, linking Stat1 activity to inflammatory signaling and immune homeostasis. Dysregulated STAT1 signaling is widely used as a mechanistic handle for studying infection-driven inflammation, autoimmunity-like phenotypes in model systems, and tumor-immune interactions in preclinical research.

Stat1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Stat1 expression without altering the underlying DNA sequence.

Stat1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Stat1 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 Stat1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Stat1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Stat1 locus and enabling the study of Stat1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Stat1 pathway restoration in tumor cells with silenced or reduced Stat1 expression.

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