TRADD CRISPR Activation Plasmid (h): sc-401177-ACT

Human TRADD (TNFRSF1A-associated via death domain) is an adaptor protein that couples activated TNF receptor 1 to downstream signaling complexes controlling inflammatory transcription and cell fate decisions. Through its death domain interactions, TRADD participates in assembly of receptor proximal complexes that regulate canonical NF-κB and MAPK pathways, and can also influence caspase-dependent apoptotic signaling depending on cellular context. TRADD-mediated signaling shapes innate immune responses, cytokine production, and stress responses, making it relevant to studies of chronic inflammation, immune dysregulation, and tumor cell survival mechanisms. Altered TNF/TRADD axis activity has been implicated in diverse disease-associated phenotypes where NF-κB-driven transcription and death receptor signaling are perturbed.

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

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

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