Transgenic zebrafish produced by retroviral infection of in vitro -cultured sperm
- 26 January 2004
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences of the United States of America
- Vol. 101 (5), 1263-1267
- https://doi.org/10.1073/pnas.0304265101
Abstract
Transgenic modification of sperm before fertilization has distinct advantages over conventional transgenic methods. The primary advantage is that the mosaicism inherent in those other techniques is avoided. A culture system using primary cultures of zebrafish male germ cells, in which the differentiation from spermatogonia to functional sperm can occur in vitro, provides the opportunity for genetic modification of sperm in vitro. Here, we report the production of transgenic zebrafish from cultured sperm. The sperm were differentiated from premeiotic germ cells infected with a pseudotyped retrovirus in vitro. The collected sperm were used to perform successful in vitro fertilizations, and transgenic embryos were identified. The transgenic fish transmitted the proviral integration to the next generation in a Mendelian fashion. We report the generation of a transgenic animal by cultured sperm and open the door to many exciting possibilities for the rapid generation of transgenic lines in model organisms such as zebrafish or other animal systems that are otherwise intractable to transgenesis.Keywords
This publication has 36 references indexed in Scilit:
- Mouse Round Spermatids Developed In Vitro from Preexisting Spermatocytes Can Produce Normal Offspring by Nuclear Injection into In Vivo-Developed Mature Oocytes1Biology of Reproduction, 2003
- Cloned zebrafish by nuclear transfer from long-term-cultured cellsNature Biotechnology, 2002
- Generation and in Vitro Differentiation of a Spermatogonial Cell LineScience, 2002
- In UteroGene Therapy: Transfer and Long-Term Expression of the BacterialneorGene in Sheep after Direct Injection of Retroviral Vectors into Preimmune FetusesHuman Gene Therapy, 1998
- Foreign Gene Expression in the Mouse Testis by Localizedin VivoGene TransferBiochemical and Biophysical Research Communications, 1997
- Development of a positive method for male stem cell-mediated gene transfer in mouse and pigMolecular Reproduction and Development, 1997
- Integration and Germ-Line Transmission of a Pseudotyped Retroviral Vector in ZebrafishScience, 1994
- Altering the Genome by Homologous RecombinationScience, 1989
- Retroviruses and insertional mutagenesis in mice: proviral integration at the Mov 34 locus leads to early embryonic death.Genes & Development, 1987
- GERM-LINE TRANSFORMATION OF MICEAnnual Review of Genetics, 1986