Mammalian Germ-Line Transgenesis by Transposition

Transposons have been used in invertebrates for transgenesis and insertional mutagens in genetic screens. We tested a functional transposon called Sleeping Beauty in the one-cell mouse embryo. In this report, we describe experiments in which transposon vectors were injected into one-cell mouse embry...

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Published inProceedings of the National Academy of Sciences - PNAS Vol. 99; no. 7; pp. 4495 - 4499
Main Authors Dupuy, Adam J., Clark, Karl, Carlson, Corey M., Fritz, Sabine, Davidson, Ann E., Markley, Karra M., Finley, Ken, Fletcher, Colin F., Ekker, Stephen C., Hackett, Perry B., Horn, Sandra, Largaespada, David A.
Format Journal Article
LanguageEnglish
Published United States National Academy of Sciences 02.04.2002
National Acad Sciences
The National Academy of Sciences
Subjects
Animals
Base Sequence
Biological Sciences
Blotting, Southern
Cloning, Molecular
DNA
DNA Transposable Elements
Embryos
Gene transfer techniques
Genes
Genetic transposition
Genetics
Genomics
Messenger RNA
Molecular Sequence Data
Mutagenesis, Insertional
Plasmids
Polymerase chain reaction
Rodents
Transgenic animals
Transposases - genetics
Transposases - physiology
Transposons
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Summary:Transposons have been used in invertebrates for transgenesis and insertional mutagens in genetic screens. We tested a functional transposon called Sleeping Beauty in the one-cell mouse embryo. In this report, we describe experiments in which transposon vectors were injected into one-cell mouse embryos with mRNA expressing the SB10 transposase enzyme. Molecular evidence of transposition was obtained by cloning of insertion sites from multiple transgenic mice produced by SB10 mRNA/transposon coinjection. We also demonstrate germ-line transmission and expression from transposed elements. This technique has promise as a germ-line transgenesis method in other vertebrate species and for insertional mutagenesis in the mouse.
Bibliography:ObjectType-Article-2
SourceType-Scholarly Journals-1
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To whom reprint requests should be addressed. E-mail: larga002@tc.umn.edu.
Edited by William F. Dove, University of Wisconsin, Madison, WI, and approved January 25, 2002
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.062630599