|
A transposon-mediated gene trap approach identifies developmentally regulated genes in zebrafish.
|
| Developmental Cell 13 July 2004 |
A transposon-mediated gene trap approach identifies developmentally regulated genes in zebrafish.
Kawakami, K., Takeda, H., Kawakami, N., Kobayashi, M., Matsuda, N., Mishina, M.
Developmental Cell, 7, 133-144, 2004.
Although zebrafish has been used as a model animal to study vertebrate development by genetic approaches, useful genetic methodologies, such as enhancer trap and gene trap, had not been developed. We developed for the first time a gene trap method in zebrafish using the Tol2 transposon system. First, we established a highly efficient transgenesis method using the transposon. When a plasmid DNA containing the Tol2 transposon vector and the transposase mRNA synthesized in vitro are co-injected into fertilized eggs, the transposon vector is inserted in the genome and transmitted to the F1 progeny at high frequencies. Then we constructed a transposon-based gene trap vector and performed a pilot screen for gene trapping. A number of fish expressing GFP in temporally and spatially restricted patterns were obtained. Endogenous transcripts are trapped by the insertions, and the insertions can interfere with expression of the trapped genes. Our gene trap approach should facilitate studies of vertebrate development and organogenesis.
 |
|
A zebrafish embryo expressing GFP in the heart constructed by the transposon-mediated gene trap method.
|
|
|
