COL4A2 CRISPR Activation Plasmid (h): sc-401462-ACT

COL4A2 encodes the alpha-2 chain of type IV collagen, a core structural component of basement membranes that assembles into collagen IV networks supporting tissue architecture and filtration barriers. By shaping extracellular matrix organization and cell–matrix adhesion, COL4A2 influences vascular integrity, epithelial polarity, and developmental morphogenesis, intersecting with pathways such as integrin/FAK signaling, ECM remodeling, and TGF-β–regulated matrix homeostasis. Perturbation of collagen IV composition can alter basement membrane mechanics and permeability, contributing to microvascular dysfunction and organ-specific pathology. Human COL4A2 variation and dysregulation have been linked to cerebrovascular and small-vessel disorders, ocular phenotypes, and kidney basement membrane abnormalities, making it a relevant target for studying ECM-driven disease mechanisms.

COL4A2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous COL4A2 expression without altering the underlying DNA sequence.

COL4A2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the COL4A2 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 COL4A2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous COL4A2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native COL4A2 locus and enabling the study of COL4A2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of COL4A2 pathway restoration in tumor cells with silenced or reduced COL4A2 expression.

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