Sox-5 CRISPR Activation Plasmid (h): sc-401854-ACT

SOX5 encodes the transcription factor Sox-5, a member of the SOX family that binds DNA through an HMG box and coordinates lineage-specific gene expression programs during development and tissue homeostasis. In human cells, Sox-5 contributes to regulation of chondrogenesis and neurogenesis by modulating enhancer activity and cooperating with other transcriptional regulators to shape differentiation, proliferation, and cell fate decisions. SOX5-associated transcriptional networks intersect with pathways controlling extracellular matrix organization and neuronal maturation, linking its activity to processes that influence tissue architecture and circuit formation. Dysregulated SOX5 expression or function has been implicated in developmental disorders and cancer-related transcriptional rewiring, supporting its value as a node for studying genotype-to-phenotype relationships.

Sox-5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SOX5 expression without altering the underlying DNA sequence.

Sox-5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SOX5 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 SOX5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Sox-5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SOX5 locus and enabling the study of Sox-5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Sox-5 pathway restoration in tumor cells with silenced or reduced SOX5 expression.

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