TRIM14 CRISPR Activation Plasmid (h2): sc-405847-ACT-2

Human TRIM14 (tripartite motif-containing 14) encodes a cytosolic TRIM family adaptor implicated in innate immune regulation, where it scaffolds signaling complexes that modulate type I interferon induction and inflammatory transcriptional programs downstream of pattern-recognition receptors. TRIM14 participates in antiviral defense pathways by influencing ubiquitin-dependent signal propagation and the stability or activity of key mediators such as cGAS–STING-, MAVS-, and NF-κB-associated components, shaping cytokine output and interferon-stimulated gene expression. Dysregulated TRIM14 signaling has been linked to altered host–pathogen responses and immune-associated pathophysiology, including chronic inflammation and contexts relevant to cancer immune evasion. TRIM14 gene editing and functional genomics tools support mechanistic studies of PRR signaling, ubiquitin pathway crosstalk, and interferon-driven transcriptional networks in human cells.

TRIM14 CRISPR Activation Plasmid (h2) provides a targeted, non-destructive approach to upregulating endogenous TRIM14 expression without altering the underlying DNA sequence.

TRIM14 CRISPR Activation Plasmid (h2) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TRIM14 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 TRIM14 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TRIM14 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TRIM14 locus and enabling the study of TRIM14-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TRIM14 pathway restoration in tumor cells with silenced or reduced TRIM14 expression.

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