TIF1γ CRISPR Activation Plasmid (h): sc-403230-ACT

TRIM33 (TIF1γ) encodes a tripartite motif-containing transcriptional coregulator that integrates chromatin-associated ubiquitin ligase activity with signal-dependent gene regulation. TIF1γ modulates SMAD-mediated TGF-β/BMP signaling, influences transcriptional elongation and chromatin remodeling, and participates in DNA damage responses through regulation of nuclear protein complexes. By tuning lineage-specific transcriptional programs and cell fate decisions, TRIM33 is studied in contexts such as hematopoietic differentiation, epithelial-to-mesenchymal transition, and stress signaling networks. Dysregulated TRIM33 function and altered TGF-β pathway output have been associated with tumor biology and other disorders involving aberrant transcriptional control.

TIF1γ CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TRIM33 expression without altering the underlying DNA sequence.

TIF1γ CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TRIM33 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 TRIM33 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TIF1γ expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TRIM33 locus and enabling the study of TIF1γ-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TIF1γ pathway restoration in tumor cells with silenced or reduced TRIM33 expression.

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