TRIM26 CRISPR Activation Plasmid (h): sc-404871-ACT

TRIM26 (tripartite motif containing 26) is a RING-type E3 ubiquitin ligase of the TRIM family that helps regulate protein stability and signaling outputs through ubiquitination-dependent mechanisms. In human cells, TRIM26 has been linked to control of innate immune and inflammatory signaling, including modulation of interferon-related pathways and downstream transcriptional programs. By influencing turnover and activity of signaling intermediates and transcription regulators, TRIM26 can affect antiviral responses, cell stress adaptation, and immune homeostasis. Dysregulated TRIM26 expression or activity has been investigated in the context of immune-mediated disorders and cancer-associated signaling networks where ubiquitin–proteasome system perturbations reshape gene expression and cell-state transitions.

TRIM26 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TRIM26 expression without altering the underlying DNA sequence.

TRIM26 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TRIM26 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 TRIM26 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TRIM26 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TRIM26 locus and enabling the study of TRIM26-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TRIM26 pathway restoration in tumor cells with silenced or reduced TRIM26 expression.

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