DcR3 CRISPR Activation Plasmid (h): sc-404980-ACT

TNFRSF6B encodes decoy receptor 3 (DcR3), a soluble member of the TNF receptor superfamily that binds Fas ligand (FASLG), LIGHT (TNFSF14), and TL1A (TNFSF15) to modulate extrinsic apoptotic signaling and inflammatory crosstalk. By sequestering these ligands, DcR3 can reshape death receptor signaling, influence NF-κB–linked immune activation programs, and affect the balance between cell survival and programmed cell death. DcR3 activity has been associated with immune evasion, altered T cell responses, and changes in cytokine-driven microenvironmental signaling in multiple disease contexts. These properties make TNFRSF6B a useful target for dissecting TNF-family ligand biology, apoptosis regulation, and inflammation-related pathways in human cell models.

DcR3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TNFRSF6B expression without altering the underlying DNA sequence.

DcR3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TNFRSF6B 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 TNFRSF6B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous DcR3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TNFRSF6B locus and enabling the study of DcR3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of DcR3 pathway restoration in tumor cells with silenced or reduced TNFRSF6B expression.

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