Emx2 CRISPR Activation Plasmid (m): sc-420170-ACT

Emx2 encodes a homeobox transcription factor that helps establish dorsal telencephalic identity and cortical arealization during mouse embryogenesis, coordinating progenitor proliferation, neuronal differentiation, and regional patterning. It acts within gene regulatory networks that integrate morphogen and growth factor cues, including interactions with WNT/β-catenin, FGF, and SHH signaling to shape forebrain development and midbrain–hindbrain boundary formation. In adult tissues, Emx2 contributes to neural stem cell dynamics and circuit specification, linking its dosage to neurodevelopmental phenotypes and altered cortical organization. Dysregulated EMX2 expression has been associated with developmental brain malformations and has been studied in the context of tumor biology and epithelial lineage programs, making it relevant for mechanistic studies of transcriptional control and cell fate.

Emx2 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Emx2 expression without altering the underlying DNA sequence.

Emx2 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Emx2 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 Emx2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Emx2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Emx2 locus and enabling the study of Emx2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Emx2 pathway restoration in tumor cells with silenced or reduced Emx2 expression.

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