TMEM173 CRISPR Activation Plasmid (m): sc-428364-ACT

Mouse Tmem173 encodes TMEM173 (STING), an endoplasmic reticulum–resident adaptor that senses cytosolic DNA indirectly through cGAMP produced by cGAS and coordinates innate immune signaling. Upon activation, TMEM173 traffics to perinuclear compartments and engages TBK1–IRF3 and NF-κB pathways to induce type I interferons, inflammatory cytokines, and antiviral restriction programs. This axis shapes antigen presentation, autophagy-linked responses, and cross-talk with cell death pathways, making Tmem173 a central node in nucleic acid–triggered inflammation. Dysregulated STING signaling has been linked to interferon-driven inflammatory phenotypes and contributes to tumor immune contexture, emphasizing its value in models of infection, autoimmunity, and cancer immunobiology.

TMEM173 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Tmem173 expression without altering the underlying DNA sequence.

TMEM173 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Tmem173 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 Tmem173 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TMEM173 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Tmem173 locus and enabling the study of TMEM173-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TMEM173 pathway restoration in tumor cells with silenced or reduced Tmem173 expression.

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