MAGE-A3 CRISPR Activation Plasmid (h): sc-402571-ACT

MAGEA3 encodes MAGE-A3, a cancer-testis antigen typically silenced in adult somatic tissues but inducible in multiple malignancies through epigenetic deregulation. MAGE-A3 is implicated in transcriptional and proteostasis control by engaging MAGE family signaling modules that can modulate E3 ubiquitin ligase activity and influence protein turnover, stress responses, and cell cycle-associated programs. Its expression is frequently linked to altered antigen presentation and immune recognition states, making it a useful molecular readout for studying tumor-associated gene regulation. As a highly restricted lineage and disease-associated marker, MAGEA3 serves as a model locus for investigating chromatin regulation, transcriptional reactivation, and downstream pathway remodeling in human cells.

MAGE-A3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MAGEA3 expression without altering the underlying DNA sequence.

MAGE-A3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MAGEA3 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 MAGEA3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MAGE-A3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MAGEA3 locus and enabling the study of MAGE-A3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MAGE-A3 pathway restoration in tumor cells with silenced or reduced MAGEA3 expression.

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