Nidogen CRISPR Activation Plasmid (h): sc-403387-ACT

Human NID1 encodes nidogen-1, a sulfated glycoprotein of basement membranes that bridges laminin and type IV collagen networks to stabilize extracellular matrix architecture. Nidogen participates in cell–matrix adhesion, tissue compartmentalization, and basement membrane–dependent regulation of migration, differentiation, and polarity, influencing signaling crosstalk with integrin/FAK and growth factor pathways. Altered nidogen abundance or organization has been associated with disrupted barrier integrity and remodeling programs observed in fibrosis, tumor invasion, and vascular or renal pathobiology. As a core basement membrane linker, NID1 is frequently examined in studies of epithelial–stromal interactions, angiogenesis, and matrix mechanics.

Nidogen CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NID1 expression without altering the underlying DNA sequence.

Nidogen CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NID1 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 NID1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Nidogen expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NID1 locus and enabling the study of Nidogen-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Nidogen pathway restoration in tumor cells with silenced or reduced NID1 expression.

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