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A Genome-Scale Resource for In Vivo Tag-Based Protein Function Exploration in C. elegans.

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

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Schanze,  Kristin
Max Planck Institute of Molecular Cell Biology and Genetics, Max Planck Society;

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Angermann,  Karolin
Max Planck Institute of Molecular Cell Biology and Genetics, Max Planck Society;

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Hasse,  Susanne
Max Planck Institute of Molecular Cell Biology and Genetics, Max Planck Society;

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Rupprecht,  Michaela
Max Planck Institute of Molecular Cell Biology and Genetics, Max Planck Society;

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Vinis,  Elisabeth
Max Planck Institute of Molecular Cell Biology and Genetics, Max Planck Society;

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Tinney,  Matthew
Max Planck Institute of Molecular Cell Biology and Genetics, Max Planck Society;

Preston,  Elicia A.
Max Planck Society;

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Zinke,  Andrea
Max Planck Institute of Molecular Cell Biology and Genetics, Max Planck Society;

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Teichgraber,  Tina
Max Planck Institute of Molecular Cell Biology and Genetics, Max Planck Society;

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Reis,  Kadri
Max Planck Institute of Molecular Cell Biology and Genetics, Max Planck Society;

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Janosch,  Stephan
Max Planck Institute of Molecular Cell Biology and Genetics, Max Planck Society;

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Ejsmont,  Radoslaw K
Max Planck Institute of Molecular Cell Biology and Genetics, Max Planck Society;

Slightam,  Cindie
Max Planck Society;

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

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Zitation

Sarov, M., Murray, J. I., Schanze, K., Pozniakovski, A., Niu, W., Angermann, K., et al. (2012). A Genome-Scale Resource for In Vivo Tag-Based Protein Function Exploration in C. elegans. Cell, 150(4), 855-866.


Zitierlink: https://hdl.handle.net/21.11116/0000-0001-07C8-6
Zusammenfassung
Understanding the in vivo dynamics of protein localization and their physical interactions is important for many problems in biology. To enable systematic protein function interrogation in a multicellular context, we built a genome-scale transgenic platform for in vivo expression of fluorescent- and affinity-tagged proteins in Caenorhabditis elegans under endogenous cis regulatory control. The platform combines computer-assisted transgene design, massively parallel DNA engineering, and next-generation sequencing to generate a resource of 14,637 genomic DNA transgenes, which covers 73% of the proteome. The multipurpose tag used allows any protein of interest to be localized in vivo or affinity purified using standard tag-based assays. We illustrate the utility of the resource by systematic chromatin immunopurification and automated 4D imaging, which produced detailed DNA binding and cell/tissue distribution maps for key transcription factor proteins.