Sox10 CRISPR Activation Plasmid (h): sc-400129-ACT

Human SOX10 encodes the transcription factor Sox10, a high-mobility group (HMG) DNA-binding protein essential for neural crest specification and lineage commitment, particularly in glial and melanocytic programs. Sox10 coordinates transcriptional networks controlling differentiation, migration, and survival, integrating with developmental pathways and chromatin remodeling to regulate target gene expression in the peripheral nervous system. Altered SOX10 activity is associated with neurocristopathies affecting enteric and peripheral glia development and with dysregulated melanocyte biology, making it a valuable node for studying cell fate decisions and gene regulatory circuitry. In biomedical research, SOX10 is frequently used to interrogate transcriptional control of myelination, pigmentation, and neural crest-derived cell plasticity.

Sox10 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SOX10 expression without altering the underlying DNA sequence.

Sox10 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SOX10 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 SOX10 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Sox10 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SOX10 locus and enabling the study of Sox10-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Sox10 pathway restoration in tumor cells with silenced or reduced SOX10 expression.

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