TRAF2 CRISPR Activation Plasmid (h): sc-400361-ACT

TRAF2 (TNF receptor–associated factor 2) is an adaptor and E3 ubiquitin ligase that couples TNF receptor superfamily members to downstream signaling, shaping inflammatory and stress responses. By coordinating K63-linked ubiquitination events, TRAF2 promotes activation of NF-κB and MAPK pathways and contributes to control of apoptosis and necroptosis through crosstalk with RIPK1-dependent complexes. TRAF2 also influences innate immune and pattern-recognition receptor signaling and can modulate JNK and noncanonical NF-κB dynamics via regulation of NIK stability. Dysregulated TRAF2 signaling has been associated with altered immune homeostasis, chronic inflammation, and oncogenic signaling programs in multiple disease contexts, making it a useful node for mechanistic studies of TNF signaling circuitry.

TRAF2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TRAF2 expression without altering the underlying DNA sequence.

TRAF2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TRAF2 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 TRAF2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TRAF2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TRAF2 locus and enabling the study of TRAF2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TRAF2 pathway restoration in tumor cells with silenced or reduced TRAF2 expression.

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