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Cell. 2012 Aug 17;150(4):855-66. doi: 10.1016/j.cell.2012.08.001.

A genome-scale resource for in vivo tag-based protein function exploration in C. elegans.

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1
TransgeneOmics Unit, Max Planck Institute of Molecular Cell Biology and Genetics, Dresden 01307, Germany. sarov@mpi-cbg.de

Abstract

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.

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PMID:
22901814
PMCID:
PMC3979301
DOI:
10.1016/j.cell.2012.08.001
[Indexed for MEDLINE]
Free PMC Article

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