MAO-A CRISPR Activation Plasmid (h): sc-401030-ACT

MAOA encodes mitochondrial outer membrane monoamine oxidase A (MAO-A), a flavin-containing enzyme that catalyzes oxidative deamination of biogenic amines such as serotonin, norepinephrine, and dopamine. By regulating monoamine turnover and generating metabolic byproducts including hydrogen peroxide, MAO-A links neurotransmitter homeostasis to redox balance and mitochondrial stress responses. MAOA activity influences synaptic signaling, neuroendocrine regulation, and inflammation-associated pathways, making it a widely studied node in neuropsychiatric and neurodegeneration research. Dysregulated MAOA expression or function has been associated with altered monoaminergic tone and behavioral phenotypes, supporting mechanistic studies in neuronal and peripheral cell models.

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

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

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