Microbubble-enhanced sonoporation: Efficient gene transduction technique for chick embryos
Microbubble-enhanced sonoporation: Efficient gene transduction technique for chick embryos
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DOI：
10.1002/gene.10232
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发表时间：
2003-10-01
期刊：
影响因子：
1.5
通讯作者：
Yamada, G
中科院分区：
文献类型：
--
作者：
Ohta, S;Suzuki, K;Yamada, G
基因转导技术是研究脊椎动物胚胎发生的基因功能的有用方法。在这项研究中，报道了一种新的基因转导技术，使用微泡增强声孔（以下简称声孔）来实现包括胚胎鸡肢芽间质在内的多个胚胎器官的异位和瞬时基因表达。本研究提出的技术具有以下优点：1）基因转导程序相对简单；2）外源基因高效转导和表达，对胚胎损伤较小。通过声孔作用，绿色荧光蛋白（GFP）或 LacZ 在肢芽间充质中错误表达，在注射部位瞬时检测到引入的表达。大多数转导的鸡胚在声孔处理后均存活，没有表现出明显的胚胎异常或细胞死亡。为了证明其评估瞬时基因转导效果的功效，Shh（音波刺猬）被转导到发育中的小鸡肢芽中。转导的肢芽显示出肢体畸形，包括部分手指重复。讨论了与该方法相关的优点和未来可能的应用。 (C) 2003 Wiley-Liss, Inc.
The gene transduction technique is a useful method to study gene functions that underlie vertebrate embryogenesis. In this study, a new gene transduction technique is reported using microbubble-enhanced sonoporation (hereafter referred to as sonoporation) to achieve ectopic and transient gene expression for several embryonic organs including embryonic chick limb bud mesenchymes. The technique proposed in this study has the advantages of 1) relatively simple gene transduction procedures, and 2) efficient exogenous gene transduction and expression with lower damages to embryos. Green fluorescent protein (GFP) or LacZ was misexpressed in limb bud mesenchymes by sonoporation, with the introduced expression transiently detected in the injected sites. Most of the transduced chick embryos survived without showing significant embryonic abnormalities or cell death after sonoporation. To demonstrate its efficacy for assessing the effect of transient gene transduction, the Shh (sonic hedgehog) was transduced into the developing chick limb bud. The transduced limb bud displayed limb malformations including partial digit duplication. Advantages and possible future applications in relation to this method are discussed. (C) 2003 Wiley-Liss, Inc.