Systems biology in human cells:- from RNAi screening via in vivo proteomics to structural biology using light microscopy
14/11/2012 - Room M203 at 14:30 - Pink Seminar
We currently do not know all human genes required for some of the most basic functions of life or how the proteins encoded by these genes work together to carry out the underlying cellular processes. To fill this gap in our knowledge, we have and are developing high throughput microscopy platforms to systematically identify genes and characterize the function and ultimately the structure of their encoded proteins in situ human cells.
For gene identification, we have integrated methods for gene silencing by RNA interference with phenotyping by fluorescence microscopy and computational image processing into a high-throughput technology platform. This enabled screening all human protein-coding genes for a number of essential cellular processes, including cell division, protein secretion and DNA repair (www.mitocheck.org). To systematically test the predicted function of the encoded proteins, we recently developed high throughput fluorescence correlation spectroscopy to quantitatively study the localization, concentration, mobility and interactions of proteins in the physiological context of the living cell in 3D over time. Finally, we are starting to take advantage of the new possibilities of super-resolution microscopy to analyze the structural organization of key protein complexes inside cells and are starting to automate those methods. Taken together, we will soon have light microscopy methods at our disposal, that allow us to carry out genetics, proteomics and structural studies in the context of the whole human cell.