Metallothionein 2A CRISPR Activation Plasmid (h): sc-410889-ACT

MT2A encodes metallothionein 2A, a cysteine-rich metal-binding protein that buffers intracellular zinc and copper, sequesters toxic heavy metals such as cadmium, and helps maintain cellular redox homeostasis. By regulating metal availability, MT2A influences metal-dependent transcriptional programs and antioxidant responses, intersecting with stress signaling and detoxification processes. MT2A expression is inducible by oxidative stress, inflammatory cues, and metal exposure, linking it to pathways that govern cell survival and adaptation to environmental insults. Dysregulated metallothionein biology has been associated with altered metal metabolism and stress tolerance observed across cancers, neurodegenerative contexts, and liver injury models, making MT2A a useful node for mechanistic studies of metal-responsive signaling.

Metallothionein 2A CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MT2A expression without altering the underlying DNA sequence.

Metallothionein 2A CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MT2A 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 MT2A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Metallothionein 2A expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MT2A locus and enabling the study of Metallothionein 2A-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Metallothionein 2A pathway restoration in tumor cells with silenced or reduced MT2A expression.

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