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Insertional mutagenesis in zebrafish rapidly identifies genes essential for early vertebrate development

Abstract

To rapidly identify genes required for early vertebrate development, we are carrying out a large-scale, insertional mutagenesis screen in zebrafish, using mouse retroviral vectors as the mutagen. We will obtain mutations in 450 to 500 different genes—roughly 20% of the genes that can be mutated to produce a visible embryonic phenotype in this species—and will clone the majority of the mutated alleles. So far, we have isolated more than 500 insertional mutants. Here we describe the first 75 insertional mutants for which the disrupted genes have been identified. In agreement with chemical mutagenesis screens, approximately one-third of the mutants have developmental defects that affect primarily one or a small number of organs, body shape or swimming behavior; the rest of the mutants show more widespread or pleiotropic abnormalities. Many of the genes we identified have not been previously assigned a biological role in vivo. Roughly 20% of the mutants result from lesions in genes for which the biochemical and cellular function of the proteins they encode cannot be deduced with confidence, if at all, from their predicted amino-acid sequences. All of the genes have either orthologs or clearly related genes in human. These results provide an unbiased view of the genetic construction kit for a vertebrate embryo, reveal the diversity of genes required for vertebrate development and suggest that hundreds of genes of unknown biochemical function essential for vertebrate development have yet to be identified.

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Figure 1: Phenotypic classification of 75 insertional zebrafish mutants.
Figure 2: Examples of mutant phenotypes isolated by insertional mutagenesis.

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Acknowledgements

We thank the following individuals for maintenance of our zebrafish colony: T. Such, S. Fuss, S. Farrington, M. Munsey and J. Whitehouse. We thank T. Angelini and A. Corcoran for raising of baby fish, J. Anderson, N. Anderson, E. Choi, J. Davenport, J. Montgomery and B. Sarkisian for assistance in screening for mutants; K. Anderson and M. Cunningham for identifying and maintaining the mutant lines of fish; B. Benfield for contributions in maintaining the laboratory and M. Lee for contributions to gene identification. We are grateful to B. Bosselman of Amgen for his continued support. This work was supported by grants from the National Center for Research Resources of the National Institutes of Health (to N.H.), Amgen (to N.H.) and the NIH (to the Center for Cancer Research at MIT). G.G. was supported by a fellowship from Amgen. A.A. was supported by a fellowship from the Ford Foundation. M.A. and E.M. were supported by fellowships from the Pew Foundation. R.N. was supported by the Anna Fuller Fund Fellowships in Molecular Oncology.

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Correspondence to Nancy Hopkins.

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Golling, G., Amsterdam, A., Sun, Z. et al. Insertional mutagenesis in zebrafish rapidly identifies genes essential for early vertebrate development. Nat Genet 31, 135–140 (2002). https://doi.org/10.1038/ng896

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