NIK CRISPR Activation Plasmid (m): sc-424749-ACT

Map3k14 encodes NF-κB–inducing kinase (NIK), a serine/threonine kinase that functions as a central regulator of the noncanonical NF-κB pathway. NIK integrates signals from select TNF receptor superfamily members to promote IKKα activation and p100 (NF-κB2) processing to p52, thereby shaping transcriptional programs that control lymphoid development, B cell survival, and inflammatory signaling. Tight control of NIK stability and activity is essential for immune homeostasis, and dysregulated NIK–NF-κB2 signaling has been linked to aberrant inflammatory states and lymphoid malignancy–associated biology in experimental models. In mouse systems, Map3k14 is widely used to dissect pathway crosstalk between noncanonical NF-κB signaling, cytokine networks, and cell fate decisions in immune and stromal compartments.

NIK CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Map3k14 expression without altering the underlying DNA sequence.

NIK CRISPR Activation Plasmid (m) 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.