MGA CRISPR Activation Plasmid (h): sc-403878-ACT

Human MGA encodes MAX gene-associated protein, a transcriptional regulator that interacts with MAX and modulates MYC/MAX network activity to shape lineage-specific gene expression programs. MGA participates in transcriptional repression and chromatin regulation, including links to non-canonical PRC1.6 complexes that constrain germ cell and developmental gene expression in somatic contexts. Through these pathways, MGA influences cell cycle control, differentiation states, and metabolic transcriptional outputs driven by E-box–dependent signaling. Disruption or altered expression of MGA has been associated with oncogenic transcriptional rewiring in multiple tumor types, making it a valuable node for mechanistic studies of MYC-related gene regulation.

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

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

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