Mnt CRISPR Activation Plasmid (h): sc-402720-ACT

Human MNT encodes Mnt, a basic helix–loop–helix leucine zipper transcription factor that heterodimerizes with MAX to bind E-box DNA elements and regulate gene expression programs controlling cell growth, metabolism, and differentiation. As a functional antagonist within the MYC/MAX/MXD network, Mnt helps restrain MYC-driven transcriptional outputs, contributing to maintenance of cellular homeostasis and checkpoint-related processes. Perturbation of MNT-regulated transcription has been linked to altered proliferative signaling and stress responses that are frequently relevant in cancer biology and lineage specification studies. Because Mnt integrates transcriptional repression and context-dependent regulation across MYC-associated pathways, it is widely studied in models of oncogenic transformation and developmental gene control.

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

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

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