Stat1 CRISPR Activation Plasmid (h): sc-400086-ACT

Human STAT1 encodes Stat1, a central transcription factor in interferon-α/β and interferon-γ signaling that is activated by JAK-mediated phosphorylation and functions downstream of cytokine receptors. Upon activation, Stat1 dimerizes and translocates to the nucleus to regulate interferon-stimulated genes controlling antiviral defense, antigen presentation, apoptosis, and cell-cycle restraint. STAT1 integrates innate and adaptive immune pathways, including crosstalk with NF-κB and IRF networks, shaping inflammatory outputs and immune surveillance. Dysregulated STAT1 activity is implicated in infection susceptibility, chronic inflammation, and oncogenic immune evasion contexts, making it a widely used node for dissecting immune signaling and transcriptional programs.

Stat1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous STAT1 expression without altering the underlying DNA sequence.

Stat1 CRISPR Activation Plasmid (h) 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.