COL4A1 CRISPR Activation Plasmid (h): sc-401792-ACT

COL4A1 encodes the α1 chain of type IV collagen, a core structural component of basement membranes that assembles into heterotrimeric networks supporting tissue architecture and barrier function. Through interactions with laminins, nidogens, perlecan, and integrins, COL4A1 influences extracellular matrix organization, mechanotransduction, and cell adhesion programs that shape vascular stability and organ development. Perturbation of COL4A1-dependent basement membrane integrity is linked to small-vessel pathology and multisystem connective tissue phenotypes, reflecting its central role in microvascular homeostasis. As a basement membrane matrix gene, COL4A1 is frequently studied in endothelial biology, fibrosis-associated remodeling, and extracellular matrix signaling pathways.

COL4A1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous COL4A1 expression without altering the underlying DNA sequence.

COL4A1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the COL4A1 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 COL4A1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous COL4A1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native COL4A1 locus and enabling the study of COL4A1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of COL4A1 pathway restoration in tumor cells with silenced or reduced COL4A1 expression.

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