TIF1α CRISPR Activation Plasmid (h): sc-403838-ACT

TRIM24 (TIF1α) is a nuclear transcriptional co-regulator that integrates chromatin cues with hormone and growth factor signaling to modulate gene expression programs controlling proliferation, differentiation, and cellular stress responses. It contains a PHD-bromodomain module that recognizes specific histone modifications, linking epigenetic state to recruitment of transcriptional machinery at promoter and enhancer regions. TRIM24 influences pathways involving nuclear receptor signaling and chromatin remodeling, and altered TRIM24 activity or expression has been associated with dysregulated transcriptional networks in cancer biology and other diseases characterized by aberrant epigenetic control. As an E3 ubiquitin ligase family member, it can also intersect with protein stability and cofactor turnover, providing a mechanistic bridge between chromatin regulation and proteostasis.

TIF1α CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TRIM24 expression without altering the underlying DNA sequence.

TIF1α CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TRIM24 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 TRIM24 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TIF1α expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TRIM24 locus and enabling the study of TIF1α-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TIF1α pathway restoration in tumor cells with silenced or reduced TRIM24 expression.

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