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BAC TransgeneOmics: a high-throughput method for exploration of protein function in mammals

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Poser,  Ina
Max Planck Institute of Molecular Cell Biology and Genetics, Max Planck Society;

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Sarov,  Mihail
Max Planck Institute of Molecular Cell Biology and Genetics, Max Planck Society;

/persons/resource/persons219747

Toyoda,  Yusuke
Max Planck Institute of Molecular Cell Biology and Genetics, Max Planck Society;

Pozniakovsky,  Andrei
Max Planck Society;

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Nitzsche,  Anja
Max Planck Institute of Molecular Cell Biology and Genetics, Max Planck Society;

/persons/resource/persons219013

Bird,  Alexander W
Max Planck Institute of Molecular Cell Biology and Genetics, Max Planck Society;

/persons/resource/persons219527

Pelletier,  Laurence
Max Planck Institute of Molecular Cell Biology and Genetics, Max Planck Society;

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Kittler,  Ralf
Max Planck Institute of Molecular Cell Biology and Genetics, Max Planck Society;

/persons/resource/persons219480

Naumann,  Ronald
Max Planck Institute of Molecular Cell Biology and Genetics, Max Planck Society;

/persons/resource/persons218985

Augsburg,  Martina
Max Planck Institute of Molecular Cell Biology and Genetics, Max Planck Society;

/persons/resource/persons219042

Buchholz,  Frank
Max Planck Institute of Molecular Cell Biology and Genetics, Max Planck Society;

/persons/resource/persons219253

Hyman,  Anthony A
Max Planck Institute of Molecular Cell Biology and Genetics, Max Planck Society;

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Citation

Poser, I., Sarov, M., Hutchins, J. R. A., Heriche, J.-K., Toyoda, Y., Pozniakovsky, A., et al. (2008). BAC TransgeneOmics: a high-throughput method for exploration of protein function in mammals. Nature Methods, 5(5), 409-415.


Cite as: https://hdl.handle.net/21.11116/0000-0001-0DE2-2
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.