NIK CRISPR Activation Plasmid (h): sc-400620-ACT

MAP3K14 encodes NF-κB–inducing kinase (NIK), a central MAP3K that drives the noncanonical NF-κB pathway by promoting IKKα-dependent processing of p100 (NFKB2) to p52 and regulating RELB transcriptional programs. NIK integrates signals downstream of receptors such as BAFFR, CD40, LTβR, and RANK to control lymphoid organogenesis, B-cell maturation, osteoclast differentiation, and inflammatory gene expression. Tight regulation of NIK stability and signaling amplitude is essential for immune homeostasis, and dysregulated MAP3K14 activity has been associated with altered NF-κB signaling in immunologic and inflammatory disorders as well as hematologic malignancy contexts. As a signaling node, NIK is frequently interrogated to map receptor-to-transcription coupling, pathway crosstalk, and adaptive stress responses.

NIK CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MAP3K14 expression without altering the underlying DNA sequence.

NIK CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MAP3K14 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 MAP3K14 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NIK expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MAP3K14 locus and enabling the study of NIK-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NIK pathway restoration in tumor cells with silenced or reduced MAP3K14 expression.

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