MAO-B CRISPR Activation Plasmid (h): sc-401732-ACT

MAOB encodes monoamine oxidase B (MAO‑B), a flavin-dependent enzyme localized to the outer mitochondrial membrane that catalyzes oxidative deamination of biogenic and dietary amines. By regulating catabolism of dopamine, phenethylamine, and related monoamines, MAO‑B influences neurotransmitter homeostasis and generates hydrogen peroxide as a byproduct, linking its activity to mitochondrial redox balance and oxidative stress pathways. MAO‑B function intersects with monoaminergic signaling, mitochondrial metabolism, and cellular stress responses in neural and peripheral tissues. Altered MAOB expression or MAO‑B activity has been associated with neurodegenerative and neuropsychiatric biology as well as inflammation- and aging-related molecular phenotypes, supporting mechanistic studies of monoamine turnover and ROS-dependent processes.

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

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

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