Thy-1/CD90 CRISPR Activation Plasmid (h): sc-400608-ACT

THY1 encodes the glycosylphosphatidylinositol-anchored surface glycoprotein Thy-1/CD90, a regulator of cell–cell and cell–matrix interactions that shapes adhesion, migration, and cytoskeletal organization. Through interactions with integrins and extracellular matrix components, Thy-1/CD90 influences focal adhesion dynamics and mechanotransduction pathways linked to cellular activation states. It is broadly used as a lineage and activation marker in stromal, endothelial, and immune contexts, and its expression is frequently studied in processes including tissue remodeling and fibrosis, neuroinflammation and neurite outgrowth, and tumor microenvironment signaling. Dysregulated THY1 expression has been associated with inflammatory and fibrotic pathobiology and with altered stromal–immune crosstalk in cancer biology.

Thy-1/CD90 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous THY1 expression without altering the underlying DNA sequence.

Thy-1/CD90 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the THY1 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 THY1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Thy-1/CD90 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native THY1 locus and enabling the study of Thy-1/CD90-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Thy-1/CD90 pathway restoration in tumor cells with silenced or reduced THY1 expression.

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