TOB1 CRISPR Activation Plasmid (h): sc-402842-ACT

Human TOB1 encodes an anti-proliferative member of the BTG/TOB family that modulates transcriptional programs controlling cell-cycle progression, differentiation, and apoptosis. TOB1 functions as a context-dependent transcriptional coregulator and interfaces with signaling networks such as TGF-β/SMAD and MAPK pathways, helping shape gene expression downstream of extracellular cues. By influencing mRNA turnover and transcriptional repression/activation complexes, TOB1 contributes to immune cell activation states and broader homeostatic control of growth. Dysregulated TOB1 expression or signaling balance has been linked to altered lymphocyte function and oncogenic processes, supporting its use as a mechanistic node in cancer biology and immunology research.

TOB1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TOB1 expression without altering the underlying DNA sequence.

TOB1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TOB1 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 TOB1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TOB1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TOB1 locus and enabling the study of TOB1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TOB1 pathway restoration in tumor cells with silenced or reduced TOB1 expression.

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