Method and probes for detecting and quantifying epigenetic regulator TET enzymatic activity. This method can be used for high throughput screening of TET inhibitors or activators.
Ten eleven translocation (TET) enzymes are important epigenetic regulators. TETs catalyze the step-wise oxidation of 5-methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC), 5-formylcytosine (5fC) and 5-carboxylcytosine (5caC) in genomic DNA, promoting DNA demethylation and gene expression changes. Assessing TET activity remains experimentally challenging.
Currently available commercial kits are indirect assays of TET activity, limited in scope, laborious, and cannot be used in vivo.
Dr. Kohli and his team at Penn have developed an improved enzyme assay for TET. Dr. Kohli has engineered specific probes for TETs that yield unique reaction intermediates, can be easily detected, and their abundance is a direct measurement of TET activity. This method presents a novel, facile, and valuable tool for evaluating TET activity.
- Measure TET enzymatic activity directly in vitro or in vivo
- High throughput screen for TET modulators, inhibitors or activators
- Direct measurement of TET enzymatic activity
- Straightforward methodology
- Can be used to detect TET1, TET2, and TET3 activity
- Can be used to determine when and where specific TET enzymes are active in particular cellular niches
Stage of Development:
In vitro data
Ghanty et al., J Am Chem Soc. 2018 Dec 19;140(50):17329
Docket # 18-8629