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A large-scale insertional mutagenesis screen in zebrafish.

A Amsterdam1, S Burgess, G Golling

  • 1Biology Department and Center for Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.

Genes & Development
|October 29, 1999
PubMed
Summary
This summary is machine-generated.

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This study details a large-scale insertional mutagenesis screen in zebrafish to identify essential developmental genes. Researchers generated thousands of mutations, aiming to clone genes crucial for vertebrate embryogenesis.

Area of Science:

  • Developmental Biology
  • Genetics
  • Zebrafish Research

Background:

  • Approximately 2500 genes are vital for zebrafish embryonic development.
  • Mutations in these essential genes cause developmental defects and embryonic lethality.
  • Large-scale mutagenesis screens are crucial for identifying these genes.

Purpose of the Study:

  • To conduct a large-scale insertional mutagenesis screen in zebrafish.
  • To isolate approximately 1000 embryonic mutations.
  • To clone a significant fraction of mutated genes essential for vertebrate embryogenesis.

Main Methods:

  • Generated ~36,000 founder zebrafish by injecting blastula-stage embryos with retroviruses.
  • Estimated 500,000-1,000,000 germline proviral insertions.

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  • Developed a protocol to breed ~80,000-150,000 insertions to homozygosity within 2 years.

    • Main Results:

    • Screened ~760 insertions, yielding 9 recessive embryonic lethal mutations.
    • Mutagenesis efficiency with the F(5) retroviral vector is lower than ENU.
    • Cloned genes from 4 recessive and 1 dominant mutation.

    Conclusions:

    • The developed protocol enables large-scale insertional mutagenesis screening in zebrafish.
    • This approach facilitates the identification and cloning of genes essential for vertebrate development.
    • Further technological improvements could make this accessible to smaller labs.