Msx-2 CRISPR Activation Plasmid (h): sc-403657-ACT

Human MSX2 encodes the homeobox transcription factor Msx-2, a nuclear regulator that integrates developmental signaling to control cell fate decisions, proliferation, and differentiation programs. Msx-2 functions downstream of BMP/TGF-β and intersects with Wnt and FGF pathways to coordinate epithelial–mesenchymal interactions, craniofacial and limb patterning, osteogenic differentiation, and tooth development. Dysregulated MSX2 activity has been linked to aberrant bone formation and craniofacial phenotypes, and altered expression is reported in contexts involving abnormal tissue remodeling and lineage commitment. As a transcriptional repressor/activator depending on cofactors, Msx-2 is frequently studied for its impact on chromatin state and downstream gene networks governing morphogenesis.

Msx-2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MSX2 expression without altering the underlying DNA sequence.

Msx-2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MSX2 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 MSX2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Msx-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MSX2 locus and enabling the study of Msx-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Msx-2 pathway restoration in tumor cells with silenced or reduced MSX2 expression.

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