Sox9 CRISPR Activation Plasmid (m): sc-423093-ACT

Mouse Sox9 encodes a high-mobility group (HMG) box transcription factor that functions as a master regulator of lineage specification and tissue morphogenesis. It integrates developmental signaling inputs, including TGF‑β/BMP, Wnt/β‑catenin, and Hedgehog pathways, to control programs involved in chondrogenesis, extracellular matrix production, and epithelial differentiation. Sox9 activity shapes cell fate decisions and progenitor maintenance through transcriptional control of genes such as Col2a1 and Acan, with context-dependent cross-talk to Runx family factors. Dysregulated Sox9 expression or function is implicated in congenital skeletal defects, fibrosis-associated remodeling, and cancer-associated lineage plasticity, making it a useful node for mechanistic studies of development and disease-related transcriptional reprogramming.

Sox9 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Sox9 expression without altering the underlying DNA sequence.

Sox9 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Sox9 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 Sox9 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Sox9 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Sox9 locus and enabling the study of Sox9-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Sox9 pathway restoration in tumor cells with silenced or reduced Sox9 expression.

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