
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
TRIM8 CRISPR Activation Plasmid (m) | sc-429995-ACT | 20 µg | $397.00 | |||
TRIM8 CRISPR Activation Plasmid (m2) | sc-429995-ACT-2 | 20 µg | $397.00 |
Trim8 encodes TRIM8, a tripartite motif (TRIM) family E3 ubiquitin ligase that regulates protein turnover and signaling output through ubiquitin-dependent mechanisms. In mouse cells, TRIM8 has been implicated in modulation of inflammatory and stress-response pathways, including crosstalk with NF-κB signaling and regulators of p53-dependent transcriptional programs, thereby influencing cell-cycle control and apoptosis. By shaping the stability and activity of pathway components, TRIM8 contributes to context-dependent control of innate immune signaling and cellular homeostasis. Dysregulated TRIM8 activity has been associated with altered immune responses and oncogenic processes in experimental models, making it a useful node for pathway and phenotype interrogation.
TRIM8 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Trim8 expression without altering the underlying DNA sequence.
TRIM8 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Trim8 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 Trim8 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TRIM8 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Trim8 locus and enabling the study of TRIM8-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TRIM8 pathway restoration in tumor cells with silenced or reduced Trim8 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.