IKK-ε CRISPR Activation Plasmid (h): sc-401786-ACT

IKBKE encodes IKK-ε, a noncanonical IκB kinase that integrates innate immune sensing with inflammatory transcriptional programs. IKK-ε phosphorylates key signaling intermediates to promote activation of IRF3/IRF7 and NF-κB-dependent gene expression, shaping type I interferon responses and cytokine production. Through crosstalk with pathways downstream of TLRs, RIG-I–like receptors, and STING, IKK-ε influences antiviral defenses, cell survival, and metabolic stress signaling. Dysregulated IKBKE activity has been linked to persistent inflammatory states and has been studied in the context of tumor-associated signaling networks and immune microenvironment remodeling.

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

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

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