COL3A1 CRISPR Activation Plasmid (h): sc-400356-ACT

COL3A1 encodes the pro-α1 chain of type III collagen, a major fibrillar extracellular matrix component enriched in extensible connective tissues such as skin, vasculature, lung, and gastrointestinal tract. It assembles into collagen fibrils that regulate matrix architecture, tensile strength, and cell–matrix signaling through integrin- and focal adhesion–linked pathways that influence adhesion, migration, and tissue remodeling. COL3A1 expression is tightly coupled to fibroblast activation programs and wound repair, and it is frequently altered in fibrotic remodeling and stromal responses within the tumor microenvironment. Dysregulation or pathogenic variation in COL3A1 is associated with heritable connective tissue disorders and contributes to mechanisms underlying vascular and organ fragility.

COL3A1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous expression without altering the underlying DNA sequence.

COL3A1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the 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 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous COL3A1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native locus and enabling the study of COL3A1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of COL3A1 pathway restoration in tumor cells with silenced or reduced expression.

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