IKK alpha CRISPR Activation Plasmid (m): sc-419658-ACT

Mouse Chuk encodes IKK alpha (IKKα), a catalytic subunit of the IκB kinase complex that couples upstream signals from TNF receptor, IL-1 receptor, Toll-like receptors, and antigen receptors to NF-κB pathway activation. Beyond canonical NF-κB signaling via IκB phosphorylation, IKKα is a key regulator of the noncanonical NF-κB cascade by promoting p100 processing to p52, influencing lymphoid organogenesis, B cell maturation, and inflammatory gene programs. IKKα also interfaces with MAPK and cell cycle-associated pathways to shape proliferation, differentiation, and stress responses. Dysregulated CHUK/IKKα activity is linked to aberrant inflammatory signaling and altered epithelial and immune homeostasis, making it relevant for mechanistic studies of immune-mediated and tumor-associated transcriptional programs.

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

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

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