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Cell ; 150(4): 855-66, 2012 Aug 17.
Article in English | MEDLINE | ID: mdl-22901814

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.


Subject(s)
Animals, Genetically Modified , Caenorhabditis elegans Proteins/analysis , Caenorhabditis elegans/genetics , Genetic Engineering/methods , Genome, Helminth , Transcription Factors/analysis , Animals , Caenorhabditis elegans/chemistry , Caenorhabditis elegans/metabolism , Caenorhabditis elegans Proteins/genetics , Transcription Factors/genetics
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