PUBLICATION

A ribozyme-mediated gene "knockdown" strategy for the identification of gene function in zebrafish

Authors
Xie, Y., Chen, X., and Wagner, T.E.
ID
ZDB-PUB-971223-1
Date
1997
Source
Proceedings of the National Academy of Sciences of the United States of America   94: 13777-13781 (Journal)
Registered Authors
Wagner, Thomas E.
Keywords
gene function determination; T7 RNA polymerase; transient expression
MeSH Terms
  • Animals
  • Base Sequence
  • DNA/genetics
  • Evaluation Studies as Topic
  • Fetal Proteins/genetics
  • Gene Expression
  • Gene Expression Regulation, Developmental
  • Gene Targeting/methods*
  • Genes, Dominant
  • Genes, Recessive
  • Microinjections
  • Phenotype
  • RNA, Catalytic/genetics*
  • RNA, Messenger/genetics
  • T-Box Domain Proteins*
  • Zebrafish/embryology
  • Zebrafish/genetics*
  • Zebrafish/metabolism
  • Zebrafish Proteins*
  • Zygote/metabolism
PubMed
9391103 Full text @ Proc. Natl. Acad. Sci. USA
Abstract
The zebrafish system offers many unique opportunities for the study of molecular biology. To date, only random mutagenesis, and not directed gene knockouts, have been demonstrated in this system. To more fully develop the potential of the zebrafish system, an approach to effectively inhibit the expression of any targeted gene in the developing zebrafish embryo has been developed. This approach uses a transient, cytoplasmic, T7 expression system, injected into the fertilized zebrafish egg to rapidly produce high levels of a ribozyme directed against the mRNA encoded by the targeted gene to inhibit its expression. In a demonstration of this strategy, expression of the recessive dominant zebrafish no tail gene was effectively inhibited by using this strategy to yield a phenotype identical to that resulting from a known defective mutation in this same gene. This, ribozyme-mediated, message deletion strategy may have use in determining the function of genetic coding sequences of unknown function.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping