GSH-2 CRISPR Activation Plasmid (h): sc-407550-ACT

Human GSX2 (GSH-2) encodes a homeobox transcription factor that regulates regional patterning and neuronal lineage specification during forebrain development, with prominent roles in ventral telencephalic progenitor identity and interneuron differentiation programs. By binding cis-regulatory elements and coordinating transcriptional networks with other developmental regulators, GSH-2 helps control proliferation-to-differentiation transitions and downstream gene expression modules that shape neural circuit formation. Dysregulated GSX2 activity or altered developmental expression has been linked to neurodevelopmental phenotypes through disruption of gene regulatory networks governing neuronal fate decisions. As a marker and functional driver of neural progenitor states, GSX2 is widely studied in stem cell differentiation models and transcriptional circuit mapping.

GSH-2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GSX2 expression without altering the underlying DNA sequence.

GSH-2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GSX2 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 GSX2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GSH-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GSX2 locus and enabling the study of GSH-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GSH-2 pathway restoration in tumor cells with silenced or reduced GSX2 expression.

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