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PMID: 14644497 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Transposon-mediated enhancer trapping in medaka.

Gene ·Vol. 322 ·2003-12-11 ·Pages 57-66

Grabher C, Henrich T, Sasado T, Arenz A, Wittbrodt J, Furutani-Seiki M

Abstract

We tested the Sleeping Beauty transposable element for its ability to efficiently insert transgenes into the genome of medaka (Oryzias latipes), an important model system for vertebrate development. We show that the SB transposon efficiently mediates integration of a reporter gene into the fish germ line. In pilot experiments, we established 174 transgenic lines with a transgenesis efficiency of 32%. Transgenes are stably transmitted to, and expressed in, subsequent generations. Interestingly, the transgenic lines show novel expression patterns with temporal and spatial specificity at a rate of 12% (21/174), likely due to both, enhancing and silencing position effects. Furthermore, promoter-dependent GFP expression in injected fish embryos is tightly correlated with germ line transmission, facilitating easy selection of founder fish. Thus, the SB transposon/transposase system provides a highly efficient tool for transgenesis in general and for the generation of novel reporter gene expression patterns in particular.

MeSH Terms
Animals Animals, Genetically Modified Base Sequence Blotting, Southern DNA/chemistry,genetics DNA Transposable Elements/genetics Embryo, Nonmammalian/metabolism Gene Expression Regulation, Developmental Green Fluorescent Proteins Luminescent Proteins/genetics,metabolism Microinjections Microscopy, Fluorescence Molecular Sequence Data Mutagenesis, Insertional/methods Oryzias/embryology,genetics Plasmids/administration & dosage,genetics Recombinant Fusion Proteins/administration & dosage,genetics,metabolism Sequence Analysis, DNA
Chemicals
DNA Transposable Elements Luminescent Proteins Recombinant Fusion Proteins Green Fluorescent Proteins DNA
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Grabher C
Developmental Biology Programme, EMBL, Meyerhofstr. 1, D-69012 Heidelberg, Germany.
Henrich T
Sasado T
Arenz A
Wittbrodt J
Furutani-Seiki M
Article Info
Journal
Gene
Abbr.
Gene
ISSN
0378-1119
Published
2003-12-11
Pages
57-66
Language
English
Region
Netherlands
NLM ID
7706761
Subset
IM
Databases
GENBANK
AJ404849
Corrections
ErratumIn
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