Sox-3 CRISPR Activation Plasmid (h): sc-403896-ACT

SOX3 encodes the Sox-3 transcription factor, a member of the SOXB1 family that binds DNA via an HMG box and regulates gene expression programs that govern neural progenitor maintenance, neuroectoderm specification, and early developmental patterning. In concert with other developmental regulators, Sox-3 modulates transcriptional networks controlling cell fate decisions, proliferation, and differentiation, including pathways that intersect with Wnt/β-catenin and Notch-dependent cues in neural tissues. Altered SOX3 dosage or regulatory disruption has been associated with neurodevelopmental phenotypes and endocrine-related traits, reflecting its role in hypothalamic–pituitary axis formation and lineage commitment. As a context-dependent transcriptional regulator, SOX3 is widely studied in pluripotent stem cell models, neural differentiation systems, and mechanistic analyses of enhancer–promoter control.

Sox-3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SOX3 expression without altering the underlying DNA sequence.

Sox-3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SOX3 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 SOX3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Sox-3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SOX3 locus and enabling the study of Sox-3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Sox-3 pathway restoration in tumor cells with silenced or reduced SOX3 expression.

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