COL14A1 CRISPR Activation Plasmid (h): sc-405019-ACT

COL14A1 encodes collagen type XIV alpha 1, a FACIT collagen that associates with fibrillar collagens in the extracellular matrix to regulate collagen fibrillogenesis, matrix organization, and tissue biomechanics. It is expressed prominently in mesenchymal-derived connective tissues and contributes to cell–matrix interactions that influence adhesion, migration, and mechanotransduction signaling. COL14A1 activity intersects with extracellular matrix remodeling processes and pathways linked to stromal organization, including collagen assembly and TGF-β–associated profibrotic responses. Dysregulated COL14A1 expression and matrix deposition patterns have been reported in fibrotic and tumor-associated stroma contexts, supporting its use as a research target in extracellular matrix biology and tissue remodeling studies.

COL14A1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous COL14A1 expression without altering the underlying DNA sequence.

COL14A1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the COL14A1 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 COL14A1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous COL14A1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native COL14A1 locus and enabling the study of COL14A1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of COL14A1 pathway restoration in tumor cells with silenced or reduced COL14A1 expression.

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