DR4 CRISPR Activation Plasmid (h): sc-400747-ACT

TNFRSF10A encodes death receptor 4 (DR4), a cell-surface member of the TNF receptor superfamily that binds TRAIL to initiate extrinsic apoptosis. Upon ligand engagement, DR4 assembles the death-inducing signaling complex, promotes caspase-8 activation, and can intersect with mitochondrial apoptosis via BID cleavage, linking receptor signaling to broader cell fate control. DR4 activity is integrated with NF-κB and MAPK pathway crosstalk that can influence inflammatory signaling and stress responses, making it a useful node for studying context-dependent survival versus death outcomes. Altered TNFRSF10A expression or pathway responsiveness has been reported across diverse cancers and immune-related settings, supporting mechanistic research into apoptotic competence, immune surveillance, and resistance phenotypes.

DR4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TNFRSF10A expression without altering the underlying DNA sequence.

DR4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TNFRSF10A 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 TNFRSF10A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous DR4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TNFRSF10A locus and enabling the study of DR4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of DR4 pathway restoration in tumor cells with silenced or reduced TNFRSF10A expression.

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