RRC ID 377
Author Kawakami K, Takeda H, Kawakami N, Kobayashi M, Matsuda N, Mishina M.
Title A transposon-mediated gene trap approach identifies developmentally regulated genes in zebrafish.
Journal Dev. Cell
Abstract We report here development of a novel gene trap method in zebrafish using the Tol2 transposon system. First, we established a highly efficient transgenesis method in which a plasmid DNA containing the Tol2 transposon vector and the transposase mRNA synthesized in vitro were coinjected into one-cell stage embryos. The transposon vector inserted in the genome could be transmitted to the F1 progeny at high frequencies, and regulated gene expression by a specific promoter could be recapitulated in transgenic fish. Then we constructed a transposon-based gene trap vector containing a splice acceptor and the GFP gene, performed a pilot screen for gene trapping, and obtained fish expressing GFP in temporally and spatially restricted patterns. We confirmed the endogenous transcripts were indeed trapped by the insertions, and the insertion could interfere with expression of the trapped gene. We propose our gene trap approach should facilitate studies of vertebrate development and organogenesis.
Volume 7(1)
Pages 133-44
Published 2004-7
DOI 10.1016/j.devcel.2004.06.005
PII S1534-5807(04)00205-9
PMID 15239961
MeSH Animals Animals, Genetically Modified Base Sequence / genetics DNA Transposable Elements / genetics* Gene Expression Regulation, Developmental / genetics* Gene Transfer Techniques Genetic Vectors / genetics Genome Green Fluorescent Proteins Luminescent Proteins / genetics Molecular Sequence Data Plasmids / genetics RNA Splicing / genetics RNA, Messenger / pharmacology Transcription, Genetic / genetics Transgenes / genetics Transposases / genetics Zebrafish / embryology* Zebrafish / genetics* Zebrafish / metabolism Zebrafish Proteins / genetics* Zebrafish Proteins / metabolism*
IF 9.616
Times Cited 466
Zebrafish ?