Stat6 CRISPR Activation Plasmid (m): sc-423180-ACT

Mouse Stat6 (signal transducer and activator of transcription 6) is a cytokine-responsive transcription factor activated downstream of IL-4 and IL-13 receptor signaling through JAK kinases, leading to Tyr phosphorylation, dimerization, and nuclear translocation. STAT6 drives gene programs associated with Th2 polarization, alternative (M2-like) macrophage activation, B cell class switching, mucus production, and tissue remodeling. Through regulation of chemokines, immunoglobulin-associated genes, and epithelial differentiation pathways, Stat6 contributes to allergic inflammation and airway hyperresponsiveness and is frequently studied in the context of asthma and atopic disease models. Its activity also intersects with tumor microenvironment immunobiology and fibrosis-related transcriptional networks, making Stat6 a useful node for pathway dissection in immunology and inflammation research.

Stat6 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Stat6 expression without altering the underlying DNA sequence.

Stat6 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Stat6 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 Stat6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Stat6 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Stat6 locus and enabling the study of Stat6-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Stat6 pathway restoration in tumor cells with silenced or reduced Stat6 expression.

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