NG2 CRISPR Activation Plasmid (h): sc-400699-ACT

CSPG4 encodes the transmembrane chondroitin sulfate proteoglycan NG2 (also known as HMW-MAA), a pericellular matrix-associated receptor that regulates cell adhesion, migration, and proliferation. NG2 participates in signaling networks linked to integrin/FAK, PDGFR, and cytoskeletal remodeling, influencing extracellular matrix interactions and cellular plasticity. In the nervous system it is a defining marker of oligodendrocyte precursor cells and modulates myelination-related processes, while in other tissues it supports perivascular and stromal cell behaviors. Dysregulated CSPG4/NG2 expression has been associated with altered invasiveness and microenvironmental interactions in multiple tumor contexts, making it a useful target for mechanistic studies of lineage state and cell–matrix signaling.

NG2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CSPG4 expression without altering the underlying DNA sequence.

NG2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CSPG4 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 CSPG4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NG2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CSPG4 locus and enabling the study of NG2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NG2 pathway restoration in tumor cells with silenced or reduced CSPG4 expression.

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