Pax-9 CRISPR Activation Plasmid (h): sc-403899-ACT

PAX9 encodes the paired box transcription factor Pax-9, a nuclear DNA-binding regulator required for embryonic patterning and organogenesis, particularly craniofacial, dental, thymic, and parathyroid development. Pax-9 modulates lineage specification programs by coordinating transcriptional networks with other developmental regulators, influencing chromatin state and downstream differentiation gene expression. Altered PAX9 dosage or function has been associated with congenital tooth agenesis and other craniofacial developmental abnormalities, making it a relevant node for studying morphogenesis and cell fate control. In adult and disease contexts, dysregulated PAX9 expression is investigated as a marker and modulator of epithelial differentiation states and transcriptional rewiring.

Pax-9 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PAX9 expression without altering the underlying DNA sequence.

Pax-9 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PAX9 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 PAX9 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Pax-9 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PAX9 locus and enabling the study of Pax-9-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Pax-9 pathway restoration in tumor cells with silenced or reduced PAX9 expression.

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