Ameloblastin CRISPR Activation Plasmid (h): sc-403086-ACT

AMBN encodes ameloblastin, a secreted extracellular matrix glycoprotein that is highly enriched during enamel formation and contributes to ameloblast adhesion, polarity, and controlled mineral deposition. Ameloblastin participates in matrix–cell signaling and coordinates enamel organ differentiation by modulating extracellular matrix organization and associated epithelial developmental programs. In addition to roles in tooth morphogenesis, AMBN expression has been linked to regulation of cell migration and differentiation in mineralized-tissue contexts, making it relevant to studies of craniofacial development and enamel structural integrity. Dysregulated AMBN activity or expression is therefore of interest for investigating molecular mechanisms underlying enamel defects and broader epithelial–matrix interactions.

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

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

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