TNF-R1 CRISPR Activation Plasmid (h): sc-400206-ACT

TNFRSF1A encodes TNF-R1, a ubiquitously expressed TNF receptor that initiates signaling in response to TNF to regulate inflammation, cell survival, and cell death. Ligand engagement promotes assembly of receptor-associated complexes that modulate NF-κB and MAPK pathways for transcriptional responses, and can also route signaling toward caspase activation and regulated necrosis depending on cellular context. Through these mechanisms, TNF-R1 shapes cytokine networks, leukocyte recruitment, and tissue homeostasis. Dysregulated TNFRSF1A/TNF-R1 signaling has been implicated in autoinflammatory phenotypes and broader inflammatory disease biology, supporting its use in mechanistic studies of immune signaling and stress responses.

TNF-R1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TNFRSF1A expression without altering the underlying DNA sequence.

TNF-R1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TNFRSF1A 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 TNFRSF1A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TNF-R1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TNFRSF1A locus and enabling the study of TNF-R1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TNF-R1 pathway restoration in tumor cells with silenced or reduced TNFRSF1A expression.

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