Beobachtung einzelner mRNA Transkripte im Zellkern und während des Kernporenkomplex vermittelten Exports in lebenden Zellen

• Applicant: Dr. David Grünwald
• Subject Area: Biochemistry ; Biophysics
• Term: from 2007 to 2010
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 52507302

The genetic information of DNA is transcribed into mRNA (messenger RNA), which gets exported into the cytoplasm. There mRNA is translated into proteins by a special machinery, called Ribosome. The control of mRNA availability is a central regulatory mechanism for cellular metabolism. Within this project single mRNA molecules shall be followed from their birth to their appearance in the cytoplasm. While transcribed, RNA gets modified and packed. RNA does exist as mRNP (messenger Ribo-Nucleo-Particle) in the nucleus. How and than these mRNPs gets modified to be recognized by the export machinery is currently not known. The time of one export event, the orientation of the mRNA during export, the question if the mRNA gets unfolded and how transport is terminated are important to elucidate our understanding of cellular function. Before it was impossible to label mRNA for microscopic applications inside living cells. Using a new viral system (MS2), it is possible to modify a gene so its mRNA contains a sequence which is highly specifically bound by the MS2 binding protein (MBP). MBP is not endogenous for mammalian cells and can be conjugated to a fluorescent protein to be used as marker for the mRNA. To track such MS2-tagged mRNAs in living cells a new multiplexing high speed microscope will be built, allowing the observation of at least three different fluorescence channels with millisecond time resolution.

• DFG Programme: Research Fellowships
• International Connection: USA
• Host: Professor Robert Singer, Ph.D.