Characterizing the movement, interactions and chemical microenvironment of a protein inside the living cell is the key towards a detailed understanding of its function. While in the past most functional studies of proteins have been performed in vitro or with cell culture experiments, it has become increasingly evident that protein function also needs to be studied on an organismal level. To achieve this objective, there is a generally acknowledged need for new and innovative approaches to study individual proteins in living animals. In this project I propose to develop a method for the covalent and specific labelling of fusion proteins in transgenic mice with synthetic probes that allows performing in vivo imaging studies of selected proteins in the living animal.
The approach is based on expression of the protein of interest as fusion with mutants of O6-alkylguanine-DNA alkyltransferase and its labelling with synthetic benzylguanine derivatives, a technology that has been developed in the host laboratory for this project and has already been applied to specifically label proteins in cell cultures. Transgenic mice will be generated using the lentivirus technique. In summary, the proposed research aims at the development of new and innovative approaches to study central biological problems and should allow establishing the covalent labelling of fusion proteins in living animals as a key technology in life sciences.
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