TNIK Gene Editing    

The serine/threonine kinase Traf2- and Nck interacting kinase (TNIK) is a member of the germinal center kinase (GCK) family. It was isolated by yeast two-hybrid screening for proteins that interact with NCK and TRAF2. TNIK was originally cloned from a human brain cDNA library, and its information level is particularly high in the brain. Genetic association studies, as well as transcription profiling of blood and postmortem brain, support a role for TNIK as a risk factor for several psychiatric diseases, including attention deficit–hyperactivity disorder, bipolar disorder, and schizophrenia. It was demonstrated that TNIK regulates Jun N-terminal kinase pathway (JNK), the actin cytoskeleton, which is an important activator of Wnt signaling and participates in the survival of many human cancer cells.

Function of TNIK

In humans, TNIK is highly expressed in the brain, heart, and skeletal muscle. TNIK has been reported to function in actin cytoskeleton and inhibiting cell spreading. In particular, the kinase domain of TNIK is necessary for cytoskeleton regulation, activation of Wnt signaling, and regulation of neurite growth. The intermediate domain of TNIK interacts with other proteins such as Nck, β-catenin, Traf2, and Nedd4-1. The GCKH domain of TNIK mediates the activation of JNK and interacts with Rap2. A loss of function mutation in TNIK led to truncated formation of TNIK protein is reported to be associated with intellectual disability in human. In an in vivo study conducted by Coba et al., TNIK knockout mice exhibited hyper locomotor behavior compared to wild type mice. This finding suggests that TNIK regulates cognitive function in mice and potential humans.

TNIK and Cancer

TNIK is one of the germinal center kinase family members involved in cytoskeleton organization and neural dendrite extension, and is reported to be related to the progression of several human cancers. TNIK is amplified or upregulated in pancreatic and gastric cancer. In particular, TNIK is a biomarker in pancreatic cancer associated with poor prognosis. In colorectal cancer (CRC), activating mutations in Wnt pathway components result in inappropriate activation of the TCF4/β-catenin transcriptional program and carcinogenesis. Approximately 80% of colorectal cancers carry a loss-of-function mutation in adenomatous polyposis coli (APC) and 5% carry activating mutations in β-catenin. Although the activating mutations in β-catenin and the loss-of-function mutation of APC were usually found to be mutually exclusive, these genetic events will lead to the constitutive activation of Wnt signaling. Notably, TNIK was considered necessary to activate Wnt signaling in colorectal cancer. In particular, it has been reported that TNIK-mediated phosphorylation of T-cell factor-4 (TCF4) at serine 154 in the nucleus results in activated transcription of Wnt target genes through the TCF4/β-catenin transcriptional complex. As a support, the TCF4/β-catenin transcription could be abrogated by inactive mutant of TNIK (K54R) with substitution at lysine 54 in the kinase domain. In addition, high expression of TNIK was significantly associated with tumor depth (T4), lymphatic invasion, and venous invasion. This result shows that elevated expression of TNIK may accelerate tumor progression and invasion.

Figure 1. TNIK is essential for colorectal cancer growth. (Yamada T, Masuda M. 2017)

TNIK Inhibitor

Wnt signaling is the main driving force of colorectal cancer. TNIK is an important regulatory component of Wnt signaling, and colorectal cancer cells are highly dependent upon the expression and catalytic activity of TNIK for proliferation. Therefore, targeting of TNIK for pharmacological intervention was anticipated to inhibit Wnt signaling and suppress the growth of colorectal cancer cells. TNIK inhibitors are currently in preclinical development. Among these inhibitors, NCB-0846 is a small-molecule TNIK inhibitor, which has been reported to have anti-Wnt activity and anti-cancer stem cell activity in colorectal cancer. NCB-0846 was designed to bind to TNIK in an inactive conformation. The function of TNIK does not seem to be limited to the regulation of Wnt signaling or colorectal carcinogenesis. Various TNIK inhibitors with different chemical structures have been developed by some pharmaceutical companies and examined in other cancers. Among these inhibitors, a CK2 (casein kinase-2)/TNIK dual inhibitor, ON108600, has been shown to target stem-like cancer cells. Tan et al. have recently reported that an anthelmintic drug, mebendazole, can selectively inhibit TNIK kinase activity. Mebendazole showed antitumor effects against a variety of cancers, and combination of mebendazole with a nonsteroidal anti-inflammatory drug reportedly reduced tumor initiation in Apc^Min/+ mice.

TNIK Gene Editing Services

CRISPR/Cas9 Platform^CB at Creative Biogene is dedicated to offering comprehensive CRISPR/Cas9 gene editing services and products for academic research, biotech research and pharmaceutical drug discovery. With deep gene editing knowledge and extensive experience in experimental operation and data processing, we help you effectively control TNIK genes knockout/knockin/point mutation in cells or animals via CRISPR/Cas9 technology.

Service	Details	Alternative cell lines or animal species
TNIK Gene Editing Cell Line Generation	gRNA design and synthesis Transfect the cell lines you're interested Select the high expression cells and sort monoclonal cell Validate the knockout/knockin/point mutation of TNIK by PCR and sequencing Provide cryogenically preserved vials of stable cells and final reports	HEK239T, Hela, HepG2, U87, Ba/F3, CHO, MDA-MB-453, MDA-MB-231NIH3T3, T47D, Neuro2a, MCF7, RKO, K562, RAW264.7, etc.
TNIK Gene Editing Animal Model Generation	TNIK gene conventional knockout animals TNIK gene conditional knockout animals TNIK point mutation animals TNIK knockin animals	Mouse, rat, rabbit, zebrafish, C. elegans, etc.

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