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Efficient gene transfer into silkworm larval tissues by a combination of sonoporation and lipofection
Author(s) -
Lee Jae Man,
Takahashi Masateru,
Mon Hiroaki,
Koga Katsumi,
Kawaguchi Yutaka,
Kusakabe Takahiro
Publication year - 2005
Publication title -
cell biology international
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.932
H-Index - 77
eISSN - 1095-8355
pISSN - 1065-6995
DOI - 10.1016/j.cellbi.2005.07.007
Subject(s) - sonoporation , malpighian tubule system , luciferase , transgene , in vivo , biology , bombyx mori , gene transfer , plasmid , microbiology and biotechnology , larva , instar , anatomy , gene , transfection , midgut , biochemistry , genetics , medicine , ultrasound , botany , microbubbles , radiology
Sonoporation (ultrasound treatment) provides a new and attractive nonviral way of in vivo gene transfer. To access the applicability of this method to the silkworm, Bombyx mori , we have compared the efficiencies of gene transfer by means of lipofection (using an appropriate agent, PDD111), sonoporation (ditto, FluoroGene™), and lipofection followed by sonoporation. By these methods, a luciferase expression plasmid was found to be markedly transferred into the haemocoel of newly ecdysed fifth instar silkworm larvae, and also into other tissues although with lower rates compared with the haemocoel. In terms of luciferase activity, the efficiencies of transgene by lipofection plus sonoporation were approximately 6 (hemocytes), 20 (silk glands), 8 (mid‐gut), 38 (fat body), 10 (Malpighian tubules), 33 (ovaries), and 16 (testes) times as high as those by lipofection or sonoporation alone. These results demonstrated that the present method is useful to introduce the exogenous DNA into insect organs in vivo.