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Genetics
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Brian Ciruna, Gilbert Weidinger, Holger Knaut, +3 , Bernard Thisse, Christine Thisse, Erez Raz, and Alexander F. Schier -3
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Abstract
We report a generally applicable strategy for transferring zygotic lethal mutations through the zebrafish germ line. By using a morpholino oligonucleotide that blocks primordial germ cell (PGC) development, we generate embryos devoid of endogenous PGCs to serve as hosts for the transplantation of germ cells derived from homozygous mutant donors. Successful transfers are identified by the localization of specifically labeled donor PGCs to the region of the developing gonad in chimeric embryos. This strategy, which results in the complete replacement of the host germ line with donor PGCs, was validated by the generation of maternal and maternal-zygotic mutants for the miles apart locus. This germ-line replacement technique provides a powerful tool for studying the maternal effects of zygotic lethal mutations. Furthermore, the ability to generate large clutches of purely mutant embryos will greatly facilitate embryological, genetic, genomic, and biochemical studies.
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Acknowledgments
We thank D. Yelon, R. Lehmann, R. G. Martinho, and members of the Schier and Yelon laboratories for discussions, and S. Zimmerman, T. Bruno, and N. Dillon for fish care. H.K. thanks Christiane Nüsslein-Volhard for support. B.C. received support from the Canadian Institutes of Health Research and the Human Frontier Science Program. H.K. was supported by a predoctoral scholarship from the Boehringer-Ingelheim Fonds. B.T. and C.T. are supported by funds from the Institut National de la Santé et de la Recherche Médicale, the Centre National de la Recherche Scientifique, the Hôpital Universitaire de Strasbourg, the Association pour la Recherche sur le Cancer, the Ligue Nationale Contre le Cancer, and the National Institutes of Health. E.R. is supported by grants from the Deutsche Forschungsgemeinschaft and the Volkswagen-Stiftung. A.F.S. is a Scholar of the McKnight Endowment Fund for Neuroscience, an Irma T. Hirschl Trust Career Scientist, and an Established Investigator of the American Heart Association, and is supported by grants from the National Institutes of Health.
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- Biological Sciences
- Genetics
Copyright
Copyright © 2002, The National Academy of Sciences.
Submission history
Received: July 31, 2002
Published online: October 23, 2002
Published in issue: November 12, 2002
Acknowledgments
We thank D. Yelon, R. Lehmann, R. G. Martinho, and members of the Schier and Yelon laboratories for discussions, and S. Zimmerman, T. Bruno, and N. Dillon for fish care. H.K. thanks Christiane Nüsslein-Volhard for support. B.C. received support from the Canadian Institutes of Health Research and the Human Frontier Science Program. H.K. was supported by a predoctoral scholarship from the Boehringer-Ingelheim Fonds. B.T. and C.T. are supported by funds from the Institut National de la Santé et de la Recherche Médicale, the Centre National de la Recherche Scientifique, the Hôpital Universitaire de Strasbourg, the Association pour la Recherche sur le Cancer, the Ligue Nationale Contre le Cancer, and the National Institutes of Health. E.R. is supported by grants from the Deutsche Forschungsgemeinschaft and the Volkswagen-Stiftung. A.F.S. is a Scholar of the McKnight Endowment Fund for Neuroscience, an Irma T. Hirschl Trust Career Scientist, and an Established Investigator of the American Heart Association, and is supported by grants from the National Institutes of Health.
Authors
Affiliations
Brian Ciruna
Developmental Genetics Program, Skirball Institute of Biomolecular Medicine, and Department of Cell Biology, New York University School of Medicine, New York, NY 10016; Max Planck Institute for Developmental Biology, Division 3, Genetics, Spemannstrasse 35/III, 72076 Tuebingen, Germany; Insitut de Genetique et de Biologie Moleculaire et Cellulaire, Centre National de la Recherche Scientifique (CNRS)/Institut National de la Santé et de la Recherche Médicale (INSERM)/Université Louis Pasteur, Centre Universitaire de Strasbourg, 67404 Illkirch Cedex, France; and Germ Cell Development, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37070 Goettingen, Germany
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Gilbert Weidinger‡
Developmental Genetics Program, Skirball Institute of Biomolecular Medicine, and Department of Cell Biology, New York University School of Medicine, New York, NY 10016; Max Planck Institute for Developmental Biology, Division 3, Genetics, Spemannstrasse 35/III, 72076 Tuebingen, Germany; Insitut de Genetique et de Biologie Moleculaire et Cellulaire, Centre National de la Recherche Scientifique (CNRS)/Institut National de la Santé et de la Recherche Médicale (INSERM)/Université Louis Pasteur, Centre Universitaire de Strasbourg, 67404 Illkirch Cedex, France; and Germ Cell Development, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37070 Goettingen, Germany
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Holger Knaut
Developmental Genetics Program, Skirball Institute of Biomolecular Medicine, and Department of Cell Biology, New York University School of Medicine, New York, NY 10016; Max Planck Institute for Developmental Biology, Division 3, Genetics, Spemannstrasse 35/III, 72076 Tuebingen, Germany; Insitut de Genetique et de Biologie Moleculaire et Cellulaire, Centre National de la Recherche Scientifique (CNRS)/Institut National de la Santé et de la Recherche Médicale (INSERM)/Université Louis Pasteur, Centre Universitaire de Strasbourg, 67404 Illkirch Cedex, France; and Germ Cell Development, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37070 Goettingen, Germany
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Bernard Thisse
Developmental Genetics Program, Skirball Institute of Biomolecular Medicine, and Department of Cell Biology, New York University School of Medicine, New York, NY 10016; Max Planck Institute for Developmental Biology, Division 3, Genetics, Spemannstrasse 35/III, 72076 Tuebingen, Germany; Insitut de Genetique et de Biologie Moleculaire et Cellulaire, Centre National de la Recherche Scientifique (CNRS)/Institut National de la Santé et de la Recherche Médicale (INSERM)/Université Louis Pasteur, Centre Universitaire de Strasbourg, 67404 Illkirch Cedex, France; and Germ Cell Development, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37070 Goettingen, Germany
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Christine Thisse
Developmental Genetics Program, Skirball Institute of Biomolecular Medicine, and Department of Cell Biology, New York University School of Medicine, New York, NY 10016; Max Planck Institute for Developmental Biology, Division 3, Genetics, Spemannstrasse 35/III, 72076 Tuebingen, Germany; Insitut de Genetique et de Biologie Moleculaire et Cellulaire, Centre National de la Recherche Scientifique (CNRS)/Institut National de la Santé et de la Recherche Médicale (INSERM)/Université Louis Pasteur, Centre Universitaire de Strasbourg, 67404 Illkirch Cedex, France; and Germ Cell Development, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37070 Goettingen, Germany
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Erez Raz
Developmental Genetics Program, Skirball Institute of Biomolecular Medicine, and Department of Cell Biology, New York University School of Medicine, New York, NY 10016; Max Planck Institute for Developmental Biology, Division 3, Genetics, Spemannstrasse 35/III, 72076 Tuebingen, Germany; Insitut de Genetique et de Biologie Moleculaire et Cellulaire, Centre National de la Recherche Scientifique (CNRS)/Institut National de la Santé et de la Recherche Médicale (INSERM)/Université Louis Pasteur, Centre Universitaire de Strasbourg, 67404 Illkirch Cedex, France; and Germ Cell Development, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37070 Goettingen, Germany
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Alexander F. Schier∥
Developmental Genetics Program, Skirball Institute of Biomolecular Medicine, and Department of Cell Biology, New York University School of Medicine, New York, NY 10016; Max Planck Institute for Developmental Biology, Division 3, Genetics, Spemannstrasse 35/III, 72076 Tuebingen, Germany; Insitut de Genetique et de Biologie Moleculaire et Cellulaire, Centre National de la Recherche Scientifique (CNRS)/Institut National de la Santé et de la Recherche Médicale (INSERM)/Université Louis Pasteur, Centre Universitaire de Strasbourg, 67404 Illkirch Cedex, France; and Germ Cell Development, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37070 Goettingen, Germany
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Notes
‡
Present address: Department of Pharmacology, University of Washington, Box 357370, Seattle, WA 98195.
∥
To whom correspondence should be addressed at: Developmental Genetics Program, Skirball Institute of Biomolecular Medicine, 4th Floor Lab 14, New York University School of Medicine, 540 First Avenue, New York, NY 10016. E-mail: [emailprotected].
Edited by Igor B. Dawid, National Institutes of Health, Bethesda, MD, and approved September 9, 2002
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B. Ciruna,G. Weidinger,H. Knaut,B. Thisse,C. Thisse,E. Raz,- &
A. F. Schier,
Production of maternal-zygotic mutant zebrafish by germ-line replacement, Proc. Natl. Acad. Sci. U.S.A. 99 (23) 14919-14924, https://doi.org/10.1073/pnas.222459999 (2002).
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Production of maternal-zygotic mutant zebrafish by germ-line replacement
Proceedings of the National Academy of Sciences
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