Thrombospondin 5 CRISPR Activation Plasmid (h): sc-402797-ACT

COMP encodes thrombospondin 5, a pentameric matricellular glycoprotein that is highly enriched in cartilage extracellular matrix and contributes to collagen fibrillogenesis, matrix assembly, and mechanotransduction in musculoskeletal tissues. Through interactions with collagens, proteoglycans, and cell-surface receptors, thrombospondin 5 influences chondrocyte adhesion and signaling pathways that coordinate endoplasmic reticulum proteostasis and extracellular matrix homeostasis. Dysregulation or pathogenic variants in COMP are associated with skeletal dysplasias and degenerative joint phenotypes, linking altered matrix organization to impaired cartilage function. As a matrix-associated regulator, COMP is widely studied in cartilage biology, connective tissue development, and extracellular matrix remodeling models.

Thrombospondin 5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous COMP expression without altering the underlying DNA sequence.

Thrombospondin 5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the COMP 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 COMP transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Thrombospondin 5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native COMP locus and enabling the study of Thrombospondin 5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Thrombospondin 5 pathway restoration in tumor cells with silenced or reduced COMP expression.

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