Generation of trangenic Xenopus laevis using the Sleeping Beauty transposon system

Using the Sleeping Beauty (SB) transposon system, we have developed a simple method for the generation of Xenopus laevis transgenic lines. The transgenesis protocol is based on the co-injection of the SB transposase mRNA and a GFP-reporter transposon into one-cell stage embryos. Transposase-dependen...

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Bibliographic Details
Published in:Transgenic research 2006-12, Vol.15 (6), p.751-760
Main Authors: SINZELLE, L, VALLIN, J, COEN, L, CHESNEAU, A, DU PASQUIER, D, POLLET, N, DEMENEIX, B, MAZABRAUD, A
Format: Article
Language:English
Subjects:
Animals
Animals, Genetically Modified
Biological and medical sciences
Biotechnology
Development Biology
DNA Transposable Elements
Embryo, Nonmammalian
Fundamental and applied biological sciences. Psychology
Gene expression
Gene Transfer Techniques
Genetic engineering
Genetic technics
Green Fluorescent Proteins
Green Fluorescent Proteins - genetics
Life Sciences
Methods. Procedures. Technologies
Microinjections
RNA, Messenger
RNA, Messenger - administration & dosage
Transgenes
Transgenic animals and transgenic plants
Transposases
Transposases - administration & dosage
Transposases - genetics
Xenopus laevis
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Summary:Using the Sleeping Beauty (SB) transposon system, we have developed a simple method for the generation of Xenopus laevis transgenic lines. The transgenesis protocol is based on the co-injection of the SB transposase mRNA and a GFP-reporter transposon into one-cell stage embryos. Transposase-dependent reporter gene expression was observed in cell clones and in hemi-transgenic animals. We determined an optimal ratio of transposase mRNA versus transposon-carrying plasmid DNA that enhanced the proportion of hemi-transgenic tadpoles. The transgene is integrated into the genome and may be transmitted to the F1 offspring depending on the germline mosaicism. Although the transposase is necessary for efficient generation of transgenic Xenopus, the integration of the transgene occurred by an non-canonical transposition process. This was observed for two transgenic lines analysed. The transposon-based technique leads to a high transgenesis rate and is simple to handle. For these reasons, it could present an attractive alternative to the classical Restriction Enzyme Mediated Integration (REMI) procedure.
ISSN:0962-8819
1573-9368
DOI:10.1007/s11248-006-9014-6