Efficient gene transfer into silkworm larval tissues by a combination of sonoporation and lipofection.
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.
Lee
Jae Man
JM
Laboratory of Silkworm Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, Hakozaki 6-10-1, Fukuoka 812-8581, Japan.
Takahashi
Masateru
M
Mon
Hiroaki
H
Koga
Katsumi
K
Kawaguchi
Yutaka
Y
Kusakabe
Takahiro
T
D000818:Animals; D012831:Bombyx; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D005229:Fatty Acids, Monounsaturated / Q000737:chemistry; D015870:Gene Expression; D020868:Gene Silencing; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D007313:Insecta; D008081:Liposomes / Q000737:chemistry; D008156:Luciferases / Q000378:metabolism; D010714:Phosphatidylethanolamines / Q000737:chemistry; D010957:Plasmids / Q000378:metabolism; D000644:Quaternary Ammonium Compounds / Q000737:chemistry; D034622:RNA Interference; D014465:Ultrasonics
C410262:1,2-dioleoyl-glycero-3-phosphatidyl ethanolamine; D005229:Fatty Acids, Monounsaturated; D008081:Liposomes; D010714:Phosphatidylethanolamines; D000644:Quaternary Ammonium Compounds; D004247:DNA; D008156:Luciferases; C070046:1,2-dioleoyloxy-3-(trimethylammonium)propane