TANK CRISPR Activation Plasmid (h): sc-402464-ACT

TANK (TRAF family member-associated NF-κB activator) is a cytosolic adaptor and scaffolding protein that coordinates signal transduction downstream of innate immune and inflammatory receptors. It interacts with TRAF proteins and kinases such as TBK1 and IKK complexes to modulate NF-κB and IRF-dependent transcription, influencing interferon responses, cytokine production, and cell survival programs. Through these pathway connections, TANK contributes to regulation of antiviral defense and inflammatory homeostasis, and its dysregulation is relevant to studies of immune signaling imbalance and inflammation-associated disease mechanisms. In human cells, altered TANK-dependent network activity can be leveraged to interrogate cross-talk between NF-κB, type I interferon signaling, and stress-response pathways.

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

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

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