TMEM173 CRISPR Activation Plasmid (h): sc-403148-ACT

TMEM173 encodes STING, an endoplasmic reticulum–resident adaptor that senses cytosolic DNA through cGAS-generated cyclic dinucleotides and initiates innate immune signaling. Upon activation, STING traffics to the Golgi and promotes TBK1- and IKK-dependent phosphorylation of IRF3 and NF-κB, driving transcriptional programs for type I interferons and inflammatory cytokines. This pathway integrates DNA damage responses, antimicrobial defense, and immunogenic cell death, and is tightly regulated to prevent aberrant inflammation. Dysregulated STING activity has been linked to autoinflammatory phenotypes, interferonopathies, and altered tumor–immune interactions, making TMEM173 a key node in immunology and cancer biology research.

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

TMEM173 CRISPR Activation Plasmid (h) 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.