Current techniques for studying gene function using cDNAs suffer from a lack of endogenous regulation. Therefore for this purpose we have developed, together with Frank Buchholz, a battery of vectors and techniques to use bacterial artificial chromosomes to analyze proteins function during cell division. With a significant amount of effort, we have used these techniques to tag over 1000 genes that have been implicated in cell division by RNAi screens, and make transformed cell pools. The important thing about this collection of tagged cell pools is that all tagged proteins are expressed on their own regulatory sequences. This library can be used for follow up experiments using mass spectrometry, RNAi or localization. We intend to complete the library of tagged BACs for 10,000 human genes in the near future and make them available for distribution. In combination with counter selection to introduce point mutants and make the genes RNAi resistant, this provides a very powerful way to perform structure-function studies in human cells. Such structure function studies in vivo, which are based on single molecule results in vitro are extremely powerful methods to test models for mitotic spindle assembly and will form the core of Mitosys, a new EU integrated project coordinated by Jan Michael Peters in Vienna. This will include modelers, biophysists and biochemists from all over Europe with an interest in modeling mitosis. We will work within this network to model mechanisms of mitotic spindle assembly.

Susanne Ernst
Marit Leuschner
Ina Poser
Andrei Pozniakovsky
Andrea Ssykor
Yusuke Toyoda

Associated Publications

    • Poser I, Sarov M, Hutchins JR, Hériché JK, Toyoda Y, Pozniakovsky A, Weigl D, Nitzsche A, Hegemann B, Bird AW, Pelletier L, Kittler R, Hua S, Naumann R, Augsburg M, Sykora MM, Hofemeister H, Zhang Y, Nasmyth K, White KP, Dietzel S, Mechtler K, Durbin R, Stewart AF, Peters JM, Buchholz F, Hyman AA.
      Nat Methods. 2008 May;5(5):409-15. BAC TransgeneOmics: a high-throughput method for exploration of protein function in mammals.