PUBLICATION

Use of an IRES bicistronic construct to trace expression of exogenously introduced mRNA in zebrafish embryos

Authors
Wang, X., Wan, H., Korzh, V., and Gong, Z.
ID
ZDB-PUB-001102-2
Date
2000
Source
Biotechniques   29(4): 814-8 (Journal)
Registered Authors
Gong, Zhiyuan, Korzh, Vladimir, Wang, Xukun
Keywords
none
MeSH Terms
  • Animals
  • Basic Helix-Loop-Helix Transcription Factors
  • Green Fluorescent Proteins
  • Homeodomain Proteins/genetics*
  • LIM-Homeodomain Proteins
  • Luminescent Proteins/genetics
  • Microinjections
  • Nerve Tissue Proteins/genetics*
  • Protein Biosynthesis*
  • RNA, Messenger/metabolism*
  • Transcription Factors
  • Zebrafish/embryology*
PubMed
11056813 Full text @ Biotechniques
Abstract
To understand gene function in developing vertebrate embryos, co-injection of an mRNA for a reporter protein and an mRNA for a testing factor is widely used. However, because of the mosaic segregation of injected nucleic acids during early embryogenesis, whether both mRNAs are translated in the same cell remains uncertain. In the present study, we tested a new system of tracing the expression of a testing gene in zebrafish using an internal ribosomal entry site (IRES) to express two proteins from the same mRNA template, thus eliminating the problem of independent translation observed in co-injection essays. A DNA construct was made for synthesizing bicistronic mRNA for NeuroD, a neurogenic transcription factor, and the enhanced green fluorescent protein (EGFP) reporter. When the bicistronic mRNA for NeuroD and EGFP was injected into zebrafish embryos at one cell stage, all EGFP-expressing embryos showed ectopic expression of neuroD mRNA and the mRNA of its potential downstream gene, islet-1. Thus, the IRES bicistronic mRNA construct might be a more convincing means of analyzing gene function in developing zebrafish embryos.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping