Stat3 CRISPR Activation Plasmid (m): sc-423176-ACT

Signal transducer and activator of transcription 3 (Stat3) is a central transcription factor in mouse cells that relays signals from cytokine and growth factor receptors to the nucleus, most prominently downstream of JAK kinases following IL-6 family signaling. Upon activation, Stat3 regulates gene programs controlling proliferation, survival, differentiation, and inflammatory responses, integrating crosstalk with MAPK and PI3K–AKT pathways. Stat3 activity influences immune cell polarization and tissue homeostasis, and dysregulated STAT3 signaling is widely studied in inflammation, fibrosis, metabolic stress, and oncogenic transcriptional networks. In the nervous system and in stem/progenitor contexts, Stat3 also contributes to lineage decisions and stress-responsive transcription.

Stat3 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Stat3 expression without altering the underlying DNA sequence.

Stat3 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Stat3 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 Stat3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Stat3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Stat3 locus and enabling the study of Stat3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Stat3 pathway restoration in tumor cells with silenced or reduced Stat3 expression.

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