RelB CRISPR Activation Plasmid (m): sc-422643-ACT

Relb encodes RelB, a member of the NF-κB/Rel family that functions as a transcription factor central to the noncanonical NF-κB pathway. RelB commonly heterodimerizes with p52 to regulate gene programs controlling immune cell differentiation, antigen presentation, lymphoid organogenesis, and inflammatory signaling downstream of receptors such as BAFFR, CD40, and lymphotoxin-β receptor. In mouse systems, RelB activity influences dendritic cell maturation and stromal-immune crosstalk, shaping cytokine and chemokine networks that coordinate tissue homeostasis. Dysregulated RelB-dependent transcription has been implicated in aberrant immune activation and chronic inflammatory states, making Relb a useful node for pathway and transcriptional network interrogation.

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

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

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