A genome-scale resource for in vivo tag-based protein function exploration in C. elegans.
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.
TransgeneOmics Unit, Max Planck Institute of Molecular Cell Biology and Genetics, Dresden 01307, Germany. sarov@mpi-cbg.de
Mihail
M
Sarov
John I
JI
Murray
Kristin
K
Schanze
Andrei
A
Pozniakovski
Wei
W
Niu
Karolin
K
Angermann
Susanne
S
Hasse
Michaela
M
Rupprecht
Elisabeth
E
Vinis
Matthew
M
Tinney
Elicia
E
Preston
Andrea
A
Zinke
Susanne
S
Enst
Tina
T
Teichgraber
Judith
J
Janette
Kadri
K
Reis
Stephan
S
Janosch
Siegfried
S
Schloissnig
Radoslaw K
RK
Ejsmont
Cindie
C
Slightam
Xiao
X
Xu
Stuart K
SK
Kim
Valerie
V
Reinke
A Francis
AF
Stewart
Michael
M
Snyder
Robert H
RH
Waterston
Anthony A
AA
Hyman
D000818:Animals; D030801:Animals, Genetically Modified; D017173:Caenorhabditis elegans / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D029742:Caenorhabditis elegans Proteins / Q000032:analysis* / Q000235:genetics; D005818:Genetic Engineering / Q000379:methods*; D049751:Genome, Helminth; D014157:Transcription Factors / Q000032:analysis* / Q000235:genetics
D029742:Caenorhabditis elegans Proteins; D014157:Transcription Factors