MIA2 CRISPR Activation Plasmid (h): sc-418205-ACT

Human MIA2 (melanoma inhibitory activity member 2) encodes a large secreted/ER-associated protein implicated in protein trafficking and extracellular matrix-related processes, with prominent functional links to hepatic cell biology. MIA2 participates in secretory pathway homeostasis and has been associated with regulation of cellular differentiation, stress responses, and tissue remodeling programs that intersect with inflammatory and fibrotic signaling. Altered MIA2 expression has been reported across liver-centered pathologies and malignancy-relevant phenotypes, supporting its use as a molecular handle for studying tumor microenvironment interactions and secretory proteome dynamics. As a result, MIA2 is of interest for mechanistic work in secretion-dependent pathways, extracellular signaling, and disease-associated transcriptional regulation.

MIA2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MIA2 expression without altering the underlying DNA sequence.

MIA2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MIA2 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 MIA2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MIA2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MIA2 locus and enabling the study of MIA2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MIA2 pathway restoration in tumor cells with silenced or reduced MIA2 expression.

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