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

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

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Genetically Modified*
  • Caenorhabditis elegans / chemistry
  • Caenorhabditis elegans / genetics*
  • Caenorhabditis elegans / metabolism
  • Caenorhabditis elegans Proteins / analysis*
  • Caenorhabditis elegans Proteins / genetics
  • Genetic Engineering / methods*
  • Genome, Helminth*
  • Transcription Factors / analysis*
  • Transcription Factors / genetics

Substances

  • Caenorhabditis elegans Proteins
  • Transcription Factors