MIAT CRISPR Activation Plasmid (h): sc-437346-ACT

MIAT (myocardial infarction associated transcript) is a human long noncoding RNA that modulates gene expression programs by influencing chromatin-associated regulation and acting as a competing endogenous RNA to shape microRNA availability. It has been implicated in coordinating pathways linked to cell cycle control, apoptosis, epithelial–mesenchymal transition, and inflammatory signaling, thereby affecting cellular differentiation and stress responses. Dysregulated MIAT expression has been reported across multiple disease-relevant contexts, including cardiovascular remodeling and diverse cancers, where it correlates with altered proliferation, invasion, and angiogenic phenotypes. As a regulatory lncRNA, MIAT is frequently studied for its role in transcriptional and post-transcriptional network architecture and for mapping upstream regulators and downstream effector genes.

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

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

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