DR6 CRISPR Activation Plasmid (h): sc-402038-ACT

Human TNFRSF21 encodes death receptor 6 (DR6), a TNF receptor superfamily member that modulates neuronal development, immune signaling, and apoptosis through receptor-mediated pathways. DR6 can engage adaptor proteins to influence caspase activation and intersects with NF-κB and MAPK signaling, shaping cytokine-responsive transcriptional programs. In neural contexts, DR6 has been linked to axon pruning and synaptic remodeling, while in immune cells it contributes to regulation of activation and cell survival. Dysregulated TNFRSF21/DR6 signaling has been associated with inflammatory phenotypes and has been investigated in the context of neurodegenerative and oncology-related mechanisms, supporting its utility as a pathway node for functional studies.

DR6 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TNFRSF21 expression without altering the underlying DNA sequence.

DR6 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TNFRSF21 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 TNFRSF21 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous DR6 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TNFRSF21 locus and enabling the study of DR6-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of DR6 pathway restoration in tumor cells with silenced or reduced TNFRSF21 expression.

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