c-Rel CRISPR Activation Plasmid (m): sc-422641-ACT

Rel encodes c-Rel, a member of the NF-κB family of transcription factors that regulates inducible gene expression programs controlling immune cell activation, proliferation, and survival in mouse tissues. Upon stimulation by antigen receptors, Toll-like receptors, or cytokines, c-Rel contributes to canonical NF-κB signaling downstream of IKK-mediated IκB degradation, enabling nuclear transcriptional responses. c-Rel is particularly relevant to lymphocyte differentiation and inflammatory transcriptional networks, where altered activity can reshape cytokine production and immune homeostasis. Dysregulated Rel/c-Rel signaling is frequently studied in the context of autoimmune-like inflammation, infection responses, and oncogenic transcriptional states in hematopoietic lineages.

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

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

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