Tenomodulin CRISPR Activation Plasmid (h): sc-401145-ACT

TNMD encodes tenomodulin, a type II transmembrane glycoprotein enriched in tendon, ligament, and other dense connective tissues where it supports extracellular matrix homeostasis and tissue maturation. Tenomodulin is associated with tenogenic differentiation programs and contributes to regulation of cell adhesion, collagen fibrillogenesis, and remodeling processes that intersect with TGF-β–driven profibrotic signaling. Altered TNMD expression has been linked to impaired tendon healing, tendinopathy, and musculoskeletal connective-tissue degeneration, and it is frequently examined alongside angiogenesis-related pathways due to reported anti-angiogenic activity of tenomodulin domains. These properties make TNMD a useful target for studying fibroblast lineage specification, mechanotransduction-adjacent remodeling responses, and disease-relevant matrix biology in human cell models.

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

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

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