Amelogenin CRISPR Activation Plasmid (h): sc-403337-ACT

AMELY encodes amelogenin, a secreted extracellular matrix protein that contributes to enamel matrix organization during odontogenesis and supports enamel mineralization through regulation of hydroxyapatite crystal growth and matrix remodeling. Amelogenin participates in tightly coordinated epithelial–mesenchymal interactions and protease-driven processing events that shape enamel formation, linking AMELY expression to developmental programs in mineralized tissues. Variation in amelogenin-associated pathways is relevant to enamel structural integrity and is frequently leveraged in human genetics because AMELX/AMELY sequence differences provide sex-informative markers in forensic and anthropological workflows. In biomedical research, AMELY serves as a tractable model for studying transcriptional control of matrix genes, differentiation state, and biomineralization-associated gene networks.

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

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

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