Efficient gene transfer into silkworm larval tissues by a combination of sonoporation and lipofection
Efficient gene transfer into silkworm larval tissues by a combination of sonoporation and lipofection
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DOI：
10.1016/j.cellbi.2005.07.007
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发表时间：
2005-11-01
影响因子：
3.9
通讯作者：
Kusakabe, T
中科院分区：
文献类型：
--
作者：
Lee, JM;Takahashi, M;Kusakabe, T
声致穿孔（超声处理）提供了一种新的和有吸引力的体内基因转移的非病毒方式。为了获得该方法对家蚕的适用性，我们比较了通过脂质转染（使用合适的试剂PDD111）、声致孔（同上，FluoroGene（TM））和脂质转染后声致孔的基因转移效率。通过这些方法，荧光素酶表达质粒被发现显着转移到新蜕皮的五龄蚕幼虫的血腔，也进入其他组织，虽然与血腔相比，以较低的速率。在荧光素酶活性方面，脂质体转染加声致穿孔的转基因效率是单独脂质体转染或声致穿孔的转基因效率的约6倍（血细胞）、20倍（丝腺）、8倍（中肠）、38倍（脂肪体）、10倍（马氏管）、33倍（卵巢）和16倍（睾丸）。这些结果表明，本方法可用于将外源DNA导入昆虫体内器官。(c)2005年国际细胞生物学联合会。由爱思唯尔有限公司出版。保留所有权利。
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 (TM)), 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 (midgut), 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. (c) 2005 International Federation for Cell Biology. Published by Elsevier Ltd. All rights reserved.