Rubinovitz-Grossman Prize
ONE LIBRARY TO MAKE THEM ALL: STREAMLINING YEAST LIBRARY CREATION BY A SWAP-TAG STRATEGY
2Zentrum für Molekulare Biologie, Universität Heidelberg (ZMBH), Heidelberg, Germany
3Bioinformatics Unit, Biological Services Department, Weizmann Institute of Science, Rehovot, Israel
4Institut für Biochemie und Molekularbiologie, ZBMZ Universität, Freiburg, Germany
5BIOSS Centre for Biological Signalling Studies, Universität Freiburg, Freiburg, Germany
6Cell Morphogenesis and Signal Transduction, German Cancer Research Center (DKFZ), Heidelberg, Germany
The yeast Saccharomyces cerevisiae is ideally suited for systematic studies relying on collections of modified strains (libraries). Despite the significance of yeast libraries and the immense variety of available tags and regulatory elements, only a few such collections exist due to the laborious process involved in their creation. To overcome the challenge of new library creation we developed a SWAp-Tag method (SWAT), in which one parental collection can be easily and efficiently modified to give rise to an endless variety of libraries of choice. We showcase the versatility of the SWAT approach by constructing a collection of ~1800 strains carrying a SWAT-GFP module at the amino termini of endomembrane proteins, and using it to create both a seamless GFP N-tagged library and an mCherry N-tagged library. These libraries enabled us to characterize hundreds of proteins for the first time. More generally, the SWAT strategy now enables fast and effortless creation of yeast libraries, opening the door for endless new ways to systematically study cell biology.
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