MAGOH CRISPR Activation Plasmid (h): sc-402667-ACT

MAGOH (mago homolog, exon junction complex subunit) is a conserved RNA-binding protein that functions as a core component of the exon junction complex, coordinating post-transcriptional gene regulation. It participates in mRNA splicing, export, localization, and nonsense-mediated mRNA decay, thereby influencing transcriptome quality control and proteome integrity. Through these processes, MAGOH contributes to cell cycle progression and neurodevelopmental programs by shaping the expression of genes involved in proliferation and differentiation. Altered dosage or dysregulation of exon junction complex components, including MAGOH, has been linked to pathogenic mechanisms in developmental disorders and cancer-associated gene expression changes, making it a useful node for studying RNA metabolism in disease-relevant contexts.

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

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

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