SLUG CRISPR Activation Plasmid (h): sc-400389-ACT

Human SNAI2 encodes the zinc-finger transcription factor SLUG, a core regulator of epithelial–mesenchymal transition (EMT) that represses epithelial adhesion programs while promoting mesenchymal traits, motility, and cell-state plasticity. SLUG integrates signaling inputs from pathways such as TGF-β, WNT/β-catenin, NOTCH, and receptor tyrosine kinases to coordinate transcriptional networks controlling differentiation, migration, and survival under stress. Altered SNAI2 activity is associated with tumor progression phenotypes, including invasion and metastasis-like behavior, and has also been linked to therapy resistance and maintenance of stem-like subpopulations in multiple cancer contexts. Beyond oncology, SLUG contributes to developmental processes and tissue remodeling, making it a useful node for studying transcriptional control of lineage decisions and cellular transitions.

SLUG CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SNAI2 expression without altering the underlying DNA sequence.

SLUG CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SNAI2 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 SNAI2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SLUG expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SNAI2 locus and enabling the study of SLUG-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SLUG pathway restoration in tumor cells with silenced or reduced SNAI2 expression.

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