COL6A3 CRISPR Activation Plasmid (h): sc-402899-ACT

COL6A3 encodes the alpha-3 chain of type VI collagen, a key extracellular matrix component that assembles into beaded microfibrils supporting tissue architecture and mechanotransduction. COL6A3-rich matrices influence cell adhesion, migration, and survival signaling through integrin- and focal adhesion–linked pathways, shaping stromal organization and extracellular remodeling. Dysregulated COL6A3 expression and collagen VI network alterations are associated with fibrosis-like remodeling programs and changes in tumor microenvironment composition, and variants in collagen VI genes have been linked to hereditary connective tissue and neuromuscular phenotypes. As a matrix-structural gene with signaling cross-talk, COL6A3 is widely used to study ECM assembly, stromal–epithelial interactions, and remodeling responses to inflammatory or mechanical cues.

COL6A3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous COL6A3 expression without altering the underlying DNA sequence.

COL6A3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the COL6A3 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 COL6A3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous COL6A3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native COL6A3 locus and enabling the study of COL6A3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of COL6A3 pathway restoration in tumor cells with silenced or reduced COL6A3 expression.

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