PUBLICATION

Insertional mutagenesis and rapid cloning of essential genes in zebrafish

Authors
Gaiano, N., Amsterdam, A., Kawakami, K., Allende, M., Becker, T., and Hopkins, N.
ID
ZDB-PUB-961219-19
Date
1996
Source
Nature   383(6603): 829-832 (Journal)
Registered Authors
Allende, Miguel L., Amsterdam, Adam, Becker, Thomas S., Gaiano, Nicholas, Hopkins, Nancy, Kawakami, Koichi
Keywords
none
MeSH Terms
  • Amino Acid Sequence
  • Animals
  • Cloning, Molecular*
  • Embryo, Nonmammalian/abnormalities
  • Embryonic Development
  • Genetic Vectors
  • Humans
  • Leukemia Virus, Murine/genetics
  • Molecular Sequence Data
  • Mutagenesis, Insertional*
  • Pharynx/embryology
  • Proteins/genetics*
  • Proviruses/genetics
  • Sequence Homology, Nucleic Acid
  • Vesicular stomatitis Indiana virus/genetics
  • Zebrafish/embryology
  • Zebrafish/genetics*
  • Zebrafish Proteins*
  • Zinc Fingers/genetics
PubMed
8893009 Full text @ Nature
Abstract
Large-scale chemical mutagenesis screens in zebrafish have led to the isolation of thousands of lethal mutations in genes that are essential for embryonic development. However, the cloning of these mutated genes is difficult at present as it requires positional cloning methods. In Drosophila, chemical mutagenesis screens were complemented with P-element insertional mutagenesis which facilitated the cloning of many genes that had been identified by chemical lesions. To facilitate the cloning of vertebrate genes that are important during embryogenesis, we have developed an insertional mutagenesis strategy in zebrafish using a retroviral vector. Here, in a pilot screen of 217 proviral insertions, we obtained three insertional mutants with embryonic lethal phenotypes, and identified two of the disrupted genes. One of these, no arches, is essential for normal pharyngeal arch development, and is homologous to the recently characterized Drosophila zinc-finger gene, clipper, which encodes a novel type of ribonuclease. As it is easy to generate tens to hundreds of thousands of proviral transgenes in zebrafish, it should now be possible to use this screening method to mutate and then rapidly clone a large number of genes affecting vertebrate developmental and cellular processes.
Genes / Markers
Figures
Expression
Phenotype
Mutation and Transgenics
Human Disease / Model Data
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping
Errata and Notes