Author/Authors :
Mihail Sarov، نويسنده , , John I. Murray، نويسنده , , Kristin Schanze، نويسنده , , Andrei Pozniakovski، نويسنده , , Wei Niu، نويسنده , , Karolin Angermann، نويسنده , , Susanne Hasse، نويسنده , , Michaela Rupprecht، نويسنده , , Elisabeth Vinis، نويسنده , , Matthew Tinney، نويسنده , , Elicia Preston، نويسنده , , Andrea Zinke، نويسنده , , Susanne Enst، نويسنده , , Tina Teichgraber، نويسنده , , Judith Janette، نويسنده , , Kadri Reis، نويسنده , , Stephan Janosch، نويسنده , , Siegfried Schloissnig، نويسنده , , Radoslaw K. Ejsmont، نويسنده , , Cindie Slightam، نويسنده , , et al.، نويسنده ,
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.
