JIK CRISPR Activation Plasmid (h): sc-408228-ACT

TAOK3 (TAO kinase 3), also known as JIK, encodes a serine/threonine kinase within the STE20 family that functions as an upstream regulator of MAPK signaling. TAOK3 participates in stress-responsive pathways, including modulation of p38 and JNK cascades, and contributes to cytoskeletal dynamics, cell-cycle control, and apoptosis-related signaling. Through phosphorylation-dependent control of downstream kinases and scaffold proteins, it influences transcriptional responses to cellular stress and growth cues. Dysregulated TAOK3 activity or expression has been associated with altered proliferative and migratory phenotypes in cancer biology and with broader signaling perturbations relevant to neurodevelopmental and inflammatory processes.

JIK CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TAOK3 expression without altering the underlying DNA sequence.

JIK CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TAOK3 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 TAOK3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous JIK expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TAOK3 locus and enabling the study of JIK-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of JIK pathway restoration in tumor cells with silenced or reduced TAOK3 expression.

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