Sox2 CRISPR Activation Plasmid (h): sc-400050-ACT

SOX2 encodes the Sox2 transcription factor, a core regulator of pluripotency and self-renewal that cooperates with OCT4 and NANOG to maintain stem cell identity while restraining lineage commitment. In human cells, Sox2 controls gene programs involved in chromatin remodeling, neural ectoderm specification, and developmental signaling networks including WNT, SHH, BMP, FGF, and Notch. Dysregulated SOX2 expression is associated with altered differentiation states, aberrant proliferation, and cellular plasticity, and has been reported in contexts of neurodevelopmental abnormalities and tumor biology. These properties make SOX2 a widely used node for studying transcriptional circuitry, epigenetic regulation, and cell fate transitions.

Sox2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SOX2 expression without altering the underlying DNA sequence.

Sox2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SOX2 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 SOX2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Sox2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SOX2 locus and enabling the study of Sox2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Sox2 pathway restoration in tumor cells with silenced or reduced SOX2 expression.

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