title: Genomic deletion induced by Tol2 transposon excision in zebrafish.
abstract: Genomic deletions induced by imprecise excision of transposons have been used to disrupt gene functions in Drosophila. To determine the excision properties of Tol2, a popular transposon in zebrafish, we took advantage of two transgenic zebrafish lines Et(gata2a:EGFP)pku684 and Et(gata2a:EGFP)pku760, and mobilized the transposon by injecting transposase mRNA into homozygous transgenic embryos. Footprint analysis showed that the Tol2 transposons were excised in either a precise or an imprecise manner. Furthermore, we identified 1093-bp and 1253-bp genomic deletions in Et(gata2a:EGFP)pku684 founder embryos flanking the 5' end of the original Tol2 insertion site, and a 1340-bp deletion in the Et(gata2a:EGFP)pku760 founder embryos flanking the 3' end of the insertion site. The mosaic Et(gata2a:EGFP)pku684 embryos were raised to adulthood and screened for germline transmission of Tol2 excision in their F(1) progeny. On average, ∼42% of the F(1) embryos displayed loss or altered EGFP patterns, demonstrating that this transposon could be efficiently excised from the zebrafish genome in the germline. Furthermore, from 59 founders, we identified one that transmitted the 1093-bp genomic deletion to its offspring. These results suggest that imprecise Tol2 transposon excision can be used as an alternative strategy to achieve gene targeting in zebrafish.
authors.affiliation: Key Laboratory of Cell Proliferation and Differentiation of the Ministry of Education, College of Life Sciences, Peking University, Beijing 100871, PR China.
authors.forename: Peng
authors.initials: P
authors.lastname: Huang
authors.forename: Linjie
authors.initials: L
authors.lastname: Xu
authors.forename: Wei
authors.initials: W
authors.lastname: Liang
authors.forename: Chi Ian
authors.initials: CI
authors.lastname: Tam
authors.forename: Yutian
authors.initials: Y
authors.lastname: Zhang
authors.forename: Fei
authors.initials: F
authors.lastname: Qi
authors.forename: Zuoyan
authors.initials: Z
authors.lastname: Zhu
authors.forename: Shuo
authors.initials: S
authors.lastname: Lin
authors.forename: Bo
authors.initials: B
authors.lastname: Zhang
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D002872:Chromosome Deletion; D004251:DNA Transposable Elements; D017353:Gene Deletion; D018390:Gene Targeting / Q000379:methods*; D017930:Genes, Reporter; D023281:Genomics / Q000379:methods; D049452:Green Fluorescent Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D015027:Zebrafish / Q000235:genetics*
chemical_list: D004251:DNA Transposable Elements; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins