MTA3 CRISPR Activation Plasmid (h): sc-402739-ACT

Human MTA3 (metastasis-associated protein 3) is a core component of the NuRD chromatin remodeling and histone deacetylase complex, coordinating nucleosome positioning and transcriptional repression across developmental and lineage-specifying gene programs. By partnering with HDACs and other NuRD subunits, MTA3 modulates epithelial differentiation, hormone-responsive transcription, and cell-cycle–linked transcriptional states through epigenetic regulation. Dysregulated MTA3 expression or altered NuRD activity has been associated with aberrant differentiation, epithelial–mesenchymal transition–related gene expression patterns, and tumor biology in multiple tissue contexts. As a chromatin regulator, MTA3 is frequently studied for its roles in transcriptional plasticity, chromatin accessibility, and network-level control of gene expression.

MTA3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MTA3 expression without altering the underlying DNA sequence.

MTA3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MTA3 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 MTA3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MTA3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MTA3 locus and enabling the study of MTA3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MTA3 pathway restoration in tumor cells with silenced or reduced MTA3 expression.

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