@article{d33d3f4b9ee44f6498748a922241ea88,
title = "BAC TransgeneOmics: A high-throughput method for exploration of protein function in mammals",
abstract = "The interpretation of genome sequences requires reliable and standardized methods to assess protein function at high throughput. Here we describe a fast and reliable pipeline to study protein function in mammalian cells based on protein tagging in bacterial artificial chromosomes (BACs). The large size of the BAC transgenes ensures the presence of most, if not all, regulatory elements and results in expression that closely matches that of the endogenous gene. We show that BAC transgenes can be rapidly and reliably generated using 96-well-format recombineering. After stable transfection of these transgenes into human tissue culture cells or mouse embryonic stem cells, the localization, protein-protein and/or protein-DNA interactions of the tagged protein are studied using generic, tag-based assays. The same high-throughput approach will be generally applicable to other model systems.",
author = "Ina Poser and Mihail Sarov and Hutchins, {James R A} and H{\'e}rich{\'e}, {Jean Karim} and Yusuke Toyoda and Andrei Pozniakovsky and Daniela Weigl and Anja Nitzsche and Bj{\"o}rn Hegemann and Bird, {Alexander W.} and Laurence Pelletier and Ralf Kittler and Sujun Hua and Ronald Naumann and Martina Augsburg and Sykora, {Martina M.} and Helmut Hofemeister and Youming Zhang and Kim Nasmyth and White, {Kevin P.} and Steffen Dietzel and Karl Mechtler and Richard Durbin and Stewart, {A. Francis} and Peters, {Jan Michael} and Frank Buchholz and Hyman, {Anthony A.}",
note = "Funding Information: We are grateful to J. Ellenberg and Z. Maliga for stimulating discussions, to K. Neugebauer for the help in establishing the ChIP protocol, and to O. Hudecz, C. Stingl and G. Mitulovic (Institute of Molecular Pathology) and A. Ssykor, M. Biesold, D. Richter, K. Kozak and D. Drechsel (Max Planck Institute for Molecular Cell Biology and Genetics) for excellent assistance. We thank I. Cheesman for helpful discussions. This work has been supported by the 6th Framework Program of the European Union, Integrated Project {\textquoteright}MitoCheck{\textquoteright} (LSHG-CT-2004-503464), and by NGFN2 grant SMP-RNAi (01GR0402). Work in the laboratories of J.-M.P. and K.M. is supported by Boehringer Ingelheim, the GenAu Program, the Austrian Research Promotion Agency (FFG), the European Science Foundation and the Austrian Science Fund (FWF) via the EuroDynaProgram. A.F.S. received funding from the 6th Framework Program of the European Union, Integrated Project {\textquoteleft}Heroic{\textquoteright} (LSHG-CT-2005-018883). K.P.W. is supported by grant 1R01HG004428-01 from the National Human Genome Research Institute of the US National Institutes of Health. R.K. is supported by a long-term fellowship of the Human Frontier Science Program Organization. Y.T. was supported by the Uehara Memorial Foundation.",
year = "2008",
month = may,
doi = "10.1038/nmeth.1199",
language = "English (US)",
volume = "5",
pages = "409--415",
journal = "Nature methods",
issn = "1548-7091",
publisher = "Public Library of Science",
number = "5",
}