Replication, integration and stable germ-line transmission of foreign sequences injected into early zebrafish embryos
- PMID: 2852096
- DOI: 10.1242/dev.103.2.403
Replication, integration and stable germ-line transmission of foreign sequences injected into early zebrafish embryos
Abstract
To generate stable lines of transgenic fish, early zebrafish embryos were injected with high concentrations of a linear bacterial plasmid. After injection, the foreign DNA was converted into a high molecular weight form and then amplified approximately tenfold during the initial rapid cleavages characteristic of the early embryo prior to gastrulation. While most of this DNA was subsequently degraded during gastrulation, some of the foreign sequences survived the gastrula stage and could be found in most of the injected fish at 3 weeks of age. Only about 5% of fish analysed 4 months after the injection retained foreign DNA in their fins, usually at less than one copy per cell. One of these fish was also found to contain about 100 copies per cell of foreign DNA in a fraction of its germ cells. Approximately 20% of the F1 offspring from this germ-line-positive parent inherited the foreign DNA, whereas 50% of F2 progeny obtained from an identified F1 individual inherited these sequences. The 50% transmission rate in F2 progeny was as expected for a single, heterozygous genomic insert. These observations indicate that injected DNA can be integrated into the fish genome, that the resulting transgenic fish are mosaic and that some of these mosaic individuals give rise to stable lines of transgenic fish.
Similar articles
-
High-frequency germ-line transmission of plasmid DNA sequences injected into fertilized zebrafish eggs.Proc Natl Acad Sci U S A. 1991 Sep 15;88(18):7953-7. doi: 10.1073/pnas.88.18.7953. Proc Natl Acad Sci U S A. 1991. PMID: 1910170 Free PMC article.
-
Stable lines of transgenic zebrafish exhibit reproducible patterns of transgene expression.Development. 1990 Jul;109(3):577-84. doi: 10.1242/dev.109.3.577. Development. 1990. PMID: 2401211
-
Transgenesis in fish.Experientia. 1991 Sep 15;47(9):891-7. doi: 10.1007/BF01929879. Experientia. 1991. PMID: 1915772 Review.
-
Integration and germ-line transmission of a pseudotyped retroviral vector in zebrafish.Science. 1994 Jul 29;265(5172):666-9. doi: 10.1126/science.8036514. Science. 1994. PMID: 8036514
-
[Search for tissue-specific regulatory elements using Tol2 transposon as an example of evolutionary synthesis of genomics and developmental biology].Ontogenez. 2008 Mar-Apr;39(2):94-9. Ontogenez. 2008. PMID: 18669290 Review. Russian.
Cited by
-
Cross-species oncogenomics using zebrafish models of cancer.Curr Opin Genet Dev. 2015 Feb;30:73-9. doi: 10.1016/j.gde.2015.04.006. Epub 2015 Jun 9. Curr Opin Genet Dev. 2015. PMID: 26070506 Free PMC article. Review.
-
Tol2 transposon-mediated transgenesis in the Midas cichlid (Amphilophus citrinellus) - towards understanding gene function and regulatory evolution in an ecological model system for rapid phenotypic diversification.BMC Dev Biol. 2017 Nov 23;17(1):15. doi: 10.1186/s12861-017-0157-x. BMC Dev Biol. 2017. PMID: 29169323 Free PMC article.
-
Arf6 and the 5'phosphatase of Synaptojanin 1 regulate autophagy in cone photoreceptors.Inside Cell. 2016 Apr;1(2):117-133. doi: 10.1002/icl3.1044. Epub 2016 Jan 16. Inside Cell. 2016. PMID: 27123470 Free PMC article.
-
Production of germ-line chimeras in zebrafish by cell transplants from genetically pigmented to albino embryos.Proc Natl Acad Sci U S A. 1992 May 15;89(10):4519-23. doi: 10.1073/pnas.89.10.4519. Proc Natl Acad Sci U S A. 1992. PMID: 1584786 Free PMC article.
-
Transgenic analysis of Dlx regulation in fish tooth development reveals evolutionary retention of enhancer function despite organ loss.Proc Natl Acad Sci U S A. 2006 Dec 19;103(51):19390-5. doi: 10.1073/pnas.0609575103. Epub 2006 Dec 4. Proc Natl Acad Sci U S A. 2006. PMID: 17146045 Free PMC article.
Publication types
MeSH terms
Substances
Grants and funding
LinkOut - more resources
Full Text Sources
Other Literature Sources
Miscellaneous