COL17A1 CRISPR Activation Plasmid (h): sc-402747-ACT

Human COL17A1 encodes collagen type XVII alpha 1, a transmembrane hemidesmosomal collagen (BP180) that anchors basal keratinocytes to the basement membrane by connecting extracellular matrix components to intracellular keratin intermediate filaments. It supports epidermal adhesion, dermal–epidermal junction integrity, and epithelial tissue homeostasis, influencing processes such as cell–matrix signaling, mechanical stability, and wound-associated remodeling. COL17A1 function is tightly linked to hemidesmosome assembly and basement membrane organization pathways, with downstream effects on epithelial barrier maintenance. Dysregulated COL17A1 expression or altered structural integrity is associated with blistering skin phenotypes and autoantigen-related dermatologic pathology, making it relevant for studies of epithelial adhesion and skin biology.

COL17A1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous COL17A1 expression without altering the underlying DNA sequence.

COL17A1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the COL17A1 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 COL17A1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous COL17A1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native COL17A1 locus and enabling the study of COL17A1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of COL17A1 pathway restoration in tumor cells with silenced or reduced COL17A1 expression.

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