nov CRISPR Activation Plasmid (h): sc-402642-ACT

Human NOV (nephroblastoma overexpressed; CCN3) encodes the secreted matricellular protein nov, a member of the CCN family that modulates cell–matrix communication and growth factor signaling. nov influences extracellular matrix remodeling, cell adhesion and migration, and differentiation programs through interactions with integrins, heparan sulfate proteoglycans, and TGF-β, Wnt/β-catenin, and Notch-associated pathways. By shaping stromal and inflammatory microenvironments, NOV expression is frequently studied in contexts of fibrosis, tissue repair, and tumor biology. Dysregulated NOV has been reported across multiple malignancies and connective tissue disorders, supporting its utility as a mechanistic node for pathway and phenotypic studies in human cells.

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

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

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