COL4A3 CRISPR Activation Plasmid (h): sc-400903-ACT

COL4A3 encodes the α3 chain of type IV collagen, a core structural component of basement membranes that contributes to network assembly and tissue-specific filtration barriers. In the kidney glomerulus, COL4A3-containing collagen IV scaffolds support podocyte–endothelial interactions, regulate extracellular matrix organization, and influence cell adhesion and mechanotransduction signaling through integrin-linked pathways. Altered COL4A3 expression or structure disrupts basement membrane integrity and is strongly associated with hereditary nephropathies, including Alport spectrum disorders and related glomerular basement membrane pathologies. Because collagen IV remodeling shapes epithelial and endothelial microenvironments, COL4A3 is also relevant to studies of fibrosis, matrix turnover, and barrier function across multiple organs.

COL4A3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous COL4A3 expression without altering the underlying DNA sequence.

COL4A3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the COL4A3 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 COL4A3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous COL4A3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native COL4A3 locus and enabling the study of COL4A3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of COL4A3 pathway restoration in tumor cells with silenced or reduced COL4A3 expression.

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