DR5 CRISPR Activation Plasmid (h): sc-401002-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- DR5 CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- DR5 CRISPR Activation Plasmid (h) consists of three plasmids at a 1:1:1 mass ratio: a plasmid encoding the deactivated Cas9 (dCas9) nuclease (D10A and N863A) fused to the transactivation domain VP64, and a blasticidin resistance gene; a plasmid encoding the MS2-p65-HSF1 fusion protein, and a hygromycin resistance gene; a plasmid encoding a target-specific 20 nt guide RNA fused to two MS2 RNA aptamers, and a puromycin resistance gene
- The resulting SAM complex binds to a site-specific region approximately 200-250 nt upstream of the transcriptional start site and provides robust recruitment of transcription factors for highly efficient gene activation
- gRNAs encoded by DR5 CRISPR Activation Plasmid (h) and DR5 CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the TNFRSF10B transcriptional start site. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: DR5 Antibody (D-6): sc-166624
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
DR5 CRISPR Activation Plasmid (h) | sc-401002-ACT | 20 µg | $397.00 |
TNFRSF10B encodes death receptor 5 (DR5), a cell-surface TNF receptor superfamily member that binds TRAIL to initiate extrinsic apoptosis. Upon ligand engagement, DR5 promotes assembly of the death-inducing signaling complex, leading to caspase-8 activation and downstream effector caspase cascades, with crosstalk to mitochondrial apoptosis via BID cleavage. DR5 signaling intersects with NF-κB and MAPK pathways and is modulated by receptor trafficking, decoy receptors, and anti-apoptotic regulators such as cFLIP and IAP proteins. Dysregulation of TNFRSF10B expression or signaling is frequently investigated in contexts of tumor cell survival, immune-mediated cytotoxicity, and mechanisms of apoptosis resistance.
DR5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TNFRSF10B expression without altering the underlying DNA sequence.
DR5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TNFRSF10B 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 TNFRSF10B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous DR5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TNFRSF10B locus and enabling the study of DR5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of DR5 pathway restoration in tumor cells with silenced or reduced TNFRSF10B expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.