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Production of maternal-zygotic mutant zebrafish by germ-line replacement
Open accessAbstract
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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Citations
Cited by
MicroRNAs Regulate Brain Morphogenesis in Zebrafish
Giraldez, A. J., Cinalli, R. M., Glasner, M. E., Enright, A. J., Thomson, J. M., Baskerville, S., Hammond, S. M., Bartel, D. P., & Schier, A. F. (2005) · Science · doi:10.1126/science.1109020 · Cited by 1,315
Self-citationCRISPRscan: designing highly efficient sgRNAs for CRISPR-Cas9 targeting in vivo
Moreno-Mateos, M. A., Vejnar, C. E., Beaudoin, J.-D., Fernandez, J. P., Mis, E. K., Khokha, M. K., & Giraldez, A. J. (2015) · Nature Methods · doi:10.1038/nmeth.3543 · Cited by 1,290
Guidelines for morpholino use in zebrafish
Stainier, D. Y. R., Raz, E., Lawson, N. D., Ekker, S. C., Burdine, R. D., Eisen, J. S., Ingham, P. W., Schulte-Merker, S., Yelon, D., Weinstein, B. M., et al. (2017) · PLOS Genetics · doi:10.1371/journal.pgen.1007000 · Cited by 336
Self-citationDampened Hedgehog signaling but normal Wnt signaling in zebrafish without cilia
Huang, P., & Schier, A. F. (2009) · Development · doi:10.1242/dev.041343 · Cited by 210
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Metadata
- Authors
- Ciruna, B., Weidinger, G., Knaut, H., Thisse, B., Thisse, C., Raz, E., & Schier, A. F.
- Venue
- Proceedings of the National Academy of Sciences
- Published
- 2002-10-23
- Volume
- 99
- Issue
- 23
- Pages
- 14919-14924
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- OpenAlex
- W2112596275
- Cited by
- 227
- References
- 0
- FWCI
- 2.06
- Type
- journal article
- Language
- en
- Open access
- green
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- Ciruna, B., Weidinger, G., Knaut, H., Thisse, B., Thisse, C., Raz, E., & Schier, A. F. (2002). Production of maternal-zygotic mutant zebrafish by germ-line replacement. Proceedings of the National Academy of Sciences. https://doi.org/10.1073/pnas.222459999
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