TRIM29 CRISPR Activation Plasmid (h): sc-402144-ACT

TRIM29 (tripartite motif containing 29) is an RBCC/TRIM family protein implicated in regulation of DNA damage responses, epithelial differentiation, and cell stress signaling. It has been reported to modulate chromatin-associated processes and influence pathways linked to genome stability, including interactions with p53-associated networks and repair factor recruitment. TRIM29 expression is frequently dysregulated across multiple tumor types, where it has been associated with altered proliferation, invasion, and responses to genotoxic stress. These features make TRIM29 a useful node for studying transcriptional control, DNA repair pathway crosstalk, and context-dependent oncogenic or tumor-suppressive phenotypes.

TRIM29 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TRIM29 expression without altering the underlying DNA sequence.

TRIM29 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TRIM29 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 TRIM29 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TRIM29 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TRIM29 locus and enabling the study of TRIM29-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TRIM29 pathway restoration in tumor cells with silenced or reduced TRIM29 expression.

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