TREX-1 CRISPR Activation Plasmid (h): sc-403875-ACT

Human TREX1 encodes TREX-1, a 3′→5′ DNA exonuclease that degrades cytosolic single- and double-stranded DNA to maintain genome and innate immune homeostasis. By clearing aberrant DNA species generated during replication stress, DNA repair, and retroelement activity, TREX-1 limits activation of nucleic acid–sensing pathways such as cGAS–STING and downstream type I interferon signaling. Dysregulated TREX1 activity is linked to inappropriate inflammatory signaling and altered DNA damage responses, making it a key node connecting DNA metabolism with antiviral defense mechanisms. TREX-1 is therefore widely studied in contexts of interferon-driven pathology, autoimmunity-associated phenotypes, and mechanisms that shape cellular responses to endogenous and exogenous DNA.

TREX-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TREX1 expression without altering the underlying DNA sequence.

TREX-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TREX1 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 TREX1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TREX-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TREX1 locus and enabling the study of TREX-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TREX-1 pathway restoration in tumor cells with silenced or reduced TREX1 expression.

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