PUBLICATION

Translational profiling through biotinylation of tagged ribosomes in zebrafish

Authors
Housley, M.P., Reischauer, S., Dieu, M., Raes, M., Stainier, D.Y., Vanhollebeke, B.
ID
ZDB-PUB-140919-5
Date
2014
Source
Development (Cambridge, England)   141(20): 3988-93 (Journal)
Registered Authors
Housley, Michael, Reischauer, Sven, Stainier, Didier, Vanhollebeke, Benoit
Keywords
none
Datasets
GEO:GSE59355
MeSH Terms
  • Animals
  • Animals, Genetically Modified
  • Biotinylation
  • Coenzyme A Ligases/chemistry
  • Gene Expression Profiling*
  • Gene Expression Regulation, Developmental*
  • Green Fluorescent Proteins/chemistry
  • In Situ Hybridization
  • Mass Spectrometry
  • Muscle, Skeletal/pathology
  • Polyribosomes/chemistry
  • RNA, Messenger/metabolism
  • RNA-Binding Proteins/physiology*
  • Ribosomal Proteins/physiology
  • Ribosomes/metabolism
  • Transcription, Genetic*
  • Zebrafish
  • Zebrafish Proteins/physiology*
PubMed
25231762 Full text @ Development
Abstract
Heterogeneity within a population of cells of the same type is a common theme in metazoan biology. Dissecting complex developmental and physiological processes crucially relies on our ability to probe the expression profile of these cell subpopulations. Current strategies rely on cell enrichment based on sequential or simultaneous use of multiple intersecting markers starting from a heterogeneous cell suspension. The extensive tissue manipulations required to generate single-cell suspensions, as well as the complexity of the required equipment, inherently complicate these approaches. Here, we propose an alternative methodology based on a genetically encoded system in the model organism Danio rerio (zebrafish). In transgenic fish, we take advantage of the combinatorial biotin transfer system, where polysome-associated mRNAs are selectively recovered from cells expressing both a tagged ribosomal subunit, Rpl10a, and the bacterial biotin ligase BirA. We have applied this technique to skeletal muscle development and identified new genes with interesting temporal expression patterns. Through this work we have thus developed additional tools for highly specific gene expression profiling.
Genes / Markers
Figures
Show all Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping