RELA/NFκB p65 CRISPR Activation Plasmid (m): sc-422642-ACT

Rela encodes the RELA/NFκB p65 subunit, a central transcriptional regulator of canonical NF-κB signaling that controls inducible gene programs in inflammation, innate and adaptive immunity, cell survival, and stress responses. In mouse cells, p65 forms functional dimers commonly with NFKB1/p50 and translocates to the nucleus following IKK-mediated degradation of IκB inhibitors, integrating cues from TNF, IL-1, TLRs, antigen receptors, and DNA damage pathways. RELA-dependent transcription modulates cytokines, chemokines, adhesion molecules, and anti-apoptotic factors, shaping macrophage activation, lymphocyte function, and tissue homeostasis. Dysregulated NF-κB/RELA activity is associated with chronic inflammatory states, autoimmunity, and oncogenic signaling networks, making Rela a frequent target for mechanistic studies of immune regulation and stress-adaptive transcription.

RELA/NFκB p65 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Rela expression without altering the underlying DNA sequence.

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

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