In vivo electroporation:: a new frontier for gene delivery and embryology
In vivo electroporation:: a new frontier for gene delivery and embryology
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DOI：
10.1046/j.1432-0436.2002.700406.x
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发表时间：
2002-06-01
期刊：
影响因子：
2.9
通讯作者：
Ogura, T
中科院分区：
文献类型：
--
作者：
Ogura, T
发育生物学的关键技术之一是将转基因导入组织并分析其对组织形态发生和器官发生的影响。在哺乳动物中，转基因方法是一种在各种组织和器官中错误表达外源基因的方法。然而，靶向表达到某些组织完全取决于特异性启动子的可用性。因此，在胚胎发生过程中控制转基因的时空表达并不是一件容易的事情。此外，如果转基因是有毒的，胚胎发育可能会被破坏，导致在所需的发育阶段之前过早死亡。作为替代系统，非洲爪蟾和斑马鱼经常使用。在这些脊椎动物模型中，可以通过将合成RNA注射到卵中来进行基因的过表达。然而，在这些系统中的遗传技术仅限于早期发展，禁止精确分析基因对器官发生在后期阶段的影响。相比之下，鸡胚长期以来一直是一个强大而有用的模型系统，在发育生物学领域占有独特的地位。尽管转基因小鸡的试验从未成功过，但是通过在蛋壳中打开的窗口容易接近发育中的胚胎使得能够进行各种技术，例如延时电影摄影术、显微外科操作（包括小鸡/鹌鹑嵌合体）、细胞和组织的移植、New体外培养等（Bortier等人，1996; Douarin等人，1996; Selleck，1996）。除了这些实验优点之外，逆转录病毒介导的基因递送以及最近的腺病毒介导的错误表达已经常规地用于鸡胚（Leber et al.，1996; Morgan和Fekete，1996）。
One of the key techniques in developmental biology is introducing transgenes into tissues and analyzing their subsequent effects on morphogenesis and organogenesis. In mammals, the transgenic approach is a way to misexpress foreign genes in various tissues and organs. However, targeting expression to certain tissues is totally dependent on the availability of specific promoters. Hence, it is not an easy task to control transgene expression temporally and spatially during embryogenesis. Further, if the transgene is toxic, embryonic development can be disrupted, resulting in premature death before the desired stages of development. As alternative systems, Xenopus and zebrafish are used frequently. In these vertebrate models, overexpression of genes can be carried out by injecting synthetic RNAs into eggs. However, genetic techniques in these systems are limited only to early development, prohibiting the precise analysis of gene effects on organogenesis in later stages. In contrast, the chick embryo has long served as a powerful and useful model system, holding a unique position in the field of developmental biology. Although trials of transgenic chicks have never been successful, easy accessibility to the developing embryo through a window opened in an eggshell enables performance of a variety of techniques, such as time-lapse cinephotomatography, microsurgical manipulations (including chick/quail chimeras), transplantation of cells and tissues, New's in vitro culture, etc. (Bortier et al., 1996; Douarin et al., 1996; Selleck, 1996). In addition to these experimental advantages, retrovirus-mediated gene delivery, and recently, adenovirus-mediated misexpression have been employed routinely in chick embryos (Leber et al., 1996; Morgan and Fekete, 1996).