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Mammalian germ-line transgenesis by transposition
Adam J Dupuy1, Karl Clark, Corey M Carlson
1The Arnold and Mabel Beckman Center for Transposon Research, Institute of Human Genetics, Department of Genetics, Cell Biology, and Development, University of Minnesota Cancer Center, University of Minnesota, Minneapolis, MN 55455, USA.
Summary
The Sleeping Beauty transposon system was successfully tested in one-cell mouse embryos, demonstrating its potential for creating transgenic mice. This method shows promise for germ-line transgenesis and insertional mutagenesis in vertebrates.
Area of Science:
- Genetics
- Molecular Biology
- Developmental Biology
Background:
- Transposons are mobile genetic elements utilized in invertebrates for genetic manipulation.
- The Sleeping Beauty (SB) transposon system is a tool for gene transfer and genetic screening.
Purpose of the Study:
- To evaluate the efficacy of the Sleeping Beauty transposon system in one-cell mouse embryos.
- To assess the potential of SB10 transposase for germ-line transgenesis and insertional mutagenesis in mice.
Main Methods:
- Injection of transposon vectors and SB10 transposase mRNA into one-cell mouse embryos.
- Molecular cloning and analysis of transposon insertion sites in resulting transgenic mice.
- Evaluation of germ-line transmission and expression of transposed elements.
Main Results:
- Successful transposition of the Sleeping Beauty system was confirmed in mouse embryos.
- Molecular evidence of transposition was obtained by cloning insertion sites.
- Germ-line transmission and expression from transposed elements were demonstrated.
Conclusions:
- The Sleeping Beauty transposon system is a viable tool for germ-line transgenesis in mice.
- This technique holds promise for insertional mutagenesis studies in the mouse.
- The SB system may be applicable for germ-line transgenesis in other vertebrate species.