GPNMB CRISPR Activation Plasmid (h): sc-402966-ACT

GPNMB (glycoprotein non-metastatic melanoma protein B) is a type I transmembrane glycoprotein enriched in myeloid lineage cells and melanocytic populations, where it contributes to cell adhesion, migration, and tissue remodeling. It is implicated in lysosome- and endosome-associated processes and can modulate inflammatory signaling and antigen-presenting cell function through interactions with extracellular matrix and immune regulatory partners. Altered GPNMB expression has been reported across contexts including melanoma biology, neuroinflammation, fibrosis, and tumor–immune microenvironment remodeling, making it a useful marker and mechanistic node in studies of stress adaptation and cellular differentiation programs. In vitro, GPNMB regulation is often studied alongside pathways controlling macrophage polarization, osteoclast activity, and epithelial–mesenchymal–like phenotypes.

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

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

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