AI Article Synopsis
- The MET receptor is crucial for development and organ renewal, but its dysregulation can lead to cancer progression, making it a valuable target for therapies.
- Developing high-throughput assays is essential for pinpointing regulators that can shutdown MET signaling.
- The described method employs high-throughput RNA interference screening alongside a receptor internalization test and In-Cell ELISA, allowing for efficient identification of factors influencing MET ubiquitination across various cell surface proteins.
Article Abstract
The tyrosine kinase MET, a receptor for hepatocyte growth factor, is a key regulator for normal development and organ renewal via stem cell maintenance. Dysregulated MET signaling contributes to tumor progression and metastasis and is considered a potent therapeutic target for a growing number of malignancies. Toward that goal it is critical to develop high-throughput assays to identify candidate regulators for the termination of MET signaling. We describe here a rapid and efficient method for identifying cellular factors required for MET ubiquitination, which utilizes high-throughput RNA interference screening (HT-siRNA) with a receptor internalization assay and an In-Cell ELISA in a 96-well format. The assay is amenable to a large array of cell surface proteins as well as genome-wide siRNA libraries, with high signal-to-background ratio and low well-to-well variability.
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| http://dx.doi.org/10.1007/978-1-4939-2309-0_26 | DOI Listing |
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