Thrombospondin 1 CRISPR Activation Plasmid (h): sc-400436-ACT

THBS1 encodes thrombospondin 1, a secreted matricellular glycoprotein that regulates extracellular matrix remodeling, cell–matrix adhesion, and cell migration through interactions with integrins, CD36, and CD47. It modulates angiogenic balance and inflammatory signaling and is a major activator of latent TGF-β, linking THBS1 to pathways controlling fibrosis, wound repair, and immune cell recruitment. Altered THBS1 expression has been associated with dysregulated vascular remodeling and stromal responses in cancer biology, as well as fibrotic and cardiovascular disease mechanisms. These properties make THBS1 a useful node for studying microenvironmental control of tissue homeostasis and stress responses.

Thrombospondin 1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous THBS1 expression without altering the underlying DNA sequence.

Thrombospondin 1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the THBS1 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 THBS1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Thrombospondin 1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native THBS1 locus and enabling the study of Thrombospondin 1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Thrombospondin 1 pathway restoration in tumor cells with silenced or reduced THBS1 expression.

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