MAGE-C1 CRISPR Activation Plasmid (h): sc-403336-ACT

Human MAGEC1 encodes MAGE-C1, a cancer/testis antigen predominantly restricted to germ cells but frequently re-expressed in malignancies, where it can modulate tumor cell programs associated with survival and immune recognition. MAGE-C1 is part of the type I MAGE family and has been linked to ubiquitin-dependent signaling through interactions with RING-domain E3 ubiquitin ligases, influencing protein stability and stress-response pathways. Dysregulated MAGEC1 expression has been reported in hematologic and solid tumor contexts and is commonly used as a molecular marker for antigen expression and transcriptional states in tumor biology. Studying MAGE-C1 supports investigations into antigen presentation, proteostasis regulation, and transcriptional control mechanisms relevant to oncogenic phenotypes.

MAGE-C1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MAGEC1 expression without altering the underlying DNA sequence.

MAGE-C1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MAGEC1 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 MAGEC1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MAGE-C1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MAGEC1 locus and enabling the study of MAGE-C1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MAGE-C1 pathway restoration in tumor cells with silenced or reduced MAGEC1 expression.

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