Thrombospondin 2 CRISPR Activation Plasmid (h): sc-401207-ACT

THBS2 encodes thrombospondin 2, a secreted matricellular glycoprotein that modulates extracellular matrix organization, cell–matrix adhesion, and tissue remodeling. Thrombospondin 2 influences integrin- and CD36-associated signaling, regulates matrix metalloproteinase activity, and shapes angiogenic and inflammatory responses within the stromal microenvironment. Through these processes, THBS2 is commonly studied in contexts of fibrosis, wound repair, and tumor–stroma interactions where extracellular matrix dynamics control cell migration and survival. Altered THBS2 expression has been associated with aberrant vascular remodeling and extracellular matrix dysregulation relevant to multiple pathophysiological states.

Thrombospondin 2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous THBS2 expression without altering the underlying DNA sequence.

Thrombospondin 2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the THBS2 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 THBS2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Thrombospondin 2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native THBS2 locus and enabling the study of Thrombospondin 2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Thrombospondin 2 pathway restoration in tumor cells with silenced or reduced THBS2 expression.

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