title: piggyBac-mediated somatic transformation of the two-spotted cricket, Gryllus bimaculatus.
abstract: Transgenic insects have been artificially produced to study functions of interesting developmental genes, using insect transposons such as piggyBac. In the case of the cricket, however, transgenic animals have not yet been successfully artificially produced. In the present study, we examined whether the piggyBac transposon functions as a tool for gene delivery in embryos of Gryllus bimaculatus. We used either a piggyBac helper plasmid or a helper RNA synthesized in vitro as a transposase source. An excision assay revealed that the helper RNA was more effective in early Gryllus eggs to transpose a marker gene of eGFP than the helper plasmid containing the piggyBac transposase gene driven by the G. bimaculatus actin3/4 promoter. Further, only when the helper RNA was used, somatic transformation of the embryo with the eGFP gene was observed. These results suggest that the piggyBac system with the helper RNA may be effective for making transgenic crickets.
authors.affiliation: Department of Biological Science and Technology, Faculty of Engineering, The University of Tokushima, 2-1 Minami-Jyosanjima-cho, Tokushima City 770-8506, Japan.
authors.forename: Yohei
authors.initials: Y
authors.lastname: Shinmyo
authors.forename: Taro
authors.initials: T
authors.lastname: Mito
authors.forename: Takashi
authors.initials: T
authors.lastname: Matsushita
authors.forename: Isao
authors.initials: I
authors.lastname: Sarashina
authors.forename: Katsuyuki
authors.initials: K
authors.lastname: Miyawaki
authors.forename: Hideyo
authors.initials: H
authors.lastname: Ohuchi
authors.forename: Sumihare
authors.initials: S
authors.lastname: Noji
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics / Q000254:growth & development; D004251:DNA Transposable Elements; D004625:Embryo, Nonmammalian / Q000502:physiology; D006135:Gryllidae / Q000196:embryology / Q000235:genetics* / Q000254:growth & development; D009024:Morphogenesis; D010957:Plasmids; D016133:Polymerase Chain Reaction / Q000379:methods; D012313:RNA / Q000235:genetics; D014170:Transformation, Genetic
chemical_list: D004251:DNA Transposable Elements; D012313:RNA