TAB3 CRISPR Activation Plasmid (h): sc-404049-ACT

Human TAB3 (TAK1-binding protein 3) is a scaffold and regulatory component of the TAB/TAK1 signaling module that couples upstream receptor and stress cues to activation of NF-κB and MAPK pathways. By coordinating assembly of kinase complexes downstream of pattern-recognition and cytokine signaling, TAB3 helps shape transcriptional programs controlling inflammatory responses, cell survival, and stress adaptation. Dysregulated signaling through TAK1-dependent networks has been implicated in chronic inflammation and oncogenic signaling contexts, making TAB3 relevant for dissecting pathway wiring and feedback control. TAB3 expression and function are therefore commonly examined in studies of innate immune signaling, signal transduction cross-talk, and stimulus-dependent transcriptional regulation.

TAB3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TAB3 expression without altering the underlying DNA sequence.

TAB3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TAB3 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 TAB3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TAB3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TAB3 locus and enabling the study of TAB3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TAB3 pathway restoration in tumor cells with silenced or reduced TAB3 expression.

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