CTHRC1 CRISPR Activation Plasmid (h): sc-418193-ACT

Human CTHRC1 (collagen triple helix repeat containing 1) encodes a secreted extracellular matrix–associated protein that modulates tissue remodeling, cell migration, and adhesion dynamics. CTHRC1 has been linked to regulation of TGF-β signaling and noncanonical Wnt/planar cell polarity pathways, influencing cytoskeletal organization and directional motility. Altered CTHRC1 expression is frequently studied in fibrotic remodeling, wound repair, and tumor-associated stromal programs where extracellular matrix turnover and invasive behavior are dysregulated. As a microenvironmental factor, CTHRC1 is used as a marker and mechanistic node to interrogate matrix-driven signaling, epithelial–mesenchymal transition–like processes, and metastatic niche formation.

CTHRC1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CTHRC1 expression without altering the underlying DNA sequence.

CTHRC1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CTHRC1 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 CTHRC1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CTHRC1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CTHRC1 locus and enabling the study of CTHRC1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CTHRC1 pathway restoration in tumor cells with silenced or reduced CTHRC1 expression.

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