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

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• 作者：Mihail Sarov ; John?I. Murray ; Kristin Schanze ; Andrei Pozniakovski ; Wei Niu ; Karolin Angermann ; Susanne Hasse ; Michaela Rupprecht ; Elisabeth Vinis ; Matthew Tinney ; Elicia Preston ; Andrea Zinke ; Susanne Enst ; Tina Teichgraber ; Judith Janette ; Kadri Reis ; Stephan Janosch ; Siegfried Schloissnig ; Radoslaw?K. Ejsmont ; Cindie Slightam ; et al.
• 刊名：Cell
• 出版年：2012
• 出版时间：17 August, 2012
• 年：2012
• 卷：150
• 期：4
• 页码：855-866
• 全文大小：2771 K

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Summary

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.