Sox-17 CRISPR Activation Plasmid (h): sc-401192-ACT

SOX17 encodes the transcription factor Sox-17, a high-mobility group (HMG) DNA-binding protein that plays essential roles in definitive endoderm specification, embryonic patterning, and lineage commitment. In human cells, Sox-17 integrates with WNT/β-catenin, TGF-β/SMAD, and Notch-associated transcriptional programs to regulate genes controlling differentiation, epithelial organization, and developmental timing. Altered SOX17 expression or regulatory disruption has been linked to defects in endoderm-derived tissues and to cancer-relevant phenotypes, including changes in proliferation, migration, and cell state stability. As a developmental regulator, SOX17 is frequently used as a marker and functional driver in stem cell differentiation and cell fate mapping studies.

Sox-17 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SOX17 expression without altering the underlying DNA sequence.

Sox-17 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SOX17 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 SOX17 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Sox-17 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SOX17 locus and enabling the study of Sox-17-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Sox-17 pathway restoration in tumor cells with silenced or reduced SOX17 expression.

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