NFκB p50 CRISPR Activation Plasmid (h): sc-400087-ACT

NFKB1 encodes the NFκB p50 subunit, a central DNA-binding component of the NF-κB transcription factor family that regulates inducible gene expression programs in immunity and stress responses. p50 commonly forms heterodimers with RELA (p65) or homodimers that shape transcriptional outputs controlling cytokines, chemokines, adhesion molecules, and anti-apoptotic factors. NFκB p50 activity is integrated downstream of TNF receptor, IL-1/TLR, antigen receptor, and DNA damage signaling through IKK-dependent processing of p105 to p50 and nuclear translocation. Dysregulated NFκB1 signaling is implicated in chronic inflammation, immune dysregulation, oncogenic transcriptional programs, and altered responses to infection and cellular stress, making it a frequent target in pathway and gene network studies.

NFκB p50 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NFKB1 expression without altering the underlying DNA sequence.

NFκB p50 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NFKB1 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 NFKB1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NFκB p50 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NFKB1 locus and enabling the study of NFκB p50-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NFκB p50 pathway restoration in tumor cells with silenced or reduced NFKB1 expression.

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