Nidogen CRISPR Activation Plasmid (m): sc-421895-ACT

Mouse Nid1 encodes nidogen (entactin), a core basement membrane glycoprotein that bridges laminin and type IV collagen networks to stabilize extracellular matrix architecture. Nidogen supports cell–matrix adhesion, tissue compartmentalization, and signal presentation that influence integrin- and growth factor–linked pathways governing migration, polarity, and differentiation. In the nervous system, Nid1 contributes to perineuronal net and vascular basement membrane integrity, impacting synaptic stabilization and neurovascular interactions. Altered nidogen-dependent matrix organization is relevant to studies of developmental defects, fibrosis-like remodeling, and invasive behavior associated with dysregulated extracellular matrix dynamics.

Nidogen CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Nid1 expression without altering the underlying DNA sequence.

Nidogen CRISPR Activation Plasmid (m) 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.