TBK1 CRISPR Activation Plasmid (h): sc-401066-ACT

Human TBK1 (TANK-binding kinase 1) is a serine/threonine kinase that integrates innate immune and stress signaling, acting downstream of pattern-recognition receptors to drive type I interferon responses through phosphorylation of IRF3/IRF7. TBK1 also modulates NF-κB signaling and regulates selective autophagy and mitophagy via phosphorylation of autophagy receptors such as OPTN and p62/SQSTM1, linking inflammatory signaling to cellular quality control. Dysregulated TBK1 activity has been associated with chronic inflammatory phenotypes and neurodegenerative processes, and TBK1 loss-of-function variants are implicated in ALS/FTD risk through impaired autophagy and altered immune signaling. These properties make TBK1 a key node for studying cGAS–STING signaling, antiviral responses, and autophagy-associated proteostasis in human cell models.

TBK1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TBK1 expression without altering the underlying DNA sequence.

TBK1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TBK1 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 TBK1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TBK1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TBK1 locus and enabling the study of TBK1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TBK1 pathway restoration in tumor cells with silenced or reduced TBK1 expression.

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