Shimogori Tomomi, Ogawa Masaharu
RIKEN, Brain Science Institute, Saitama, Japan.
Dev Growth Differ. 2008 Aug;50(6):499-506. doi: 10.1111/j.1440-169X.2008.01045.x. Epub 2008 May 14.
Mouse genetic manipulations, such as the production of gene knock-out, knock-in, and transgenic mice, have provided excellent systems for analysis of numerous genes functioning during development. Nevertheless, the lack of specific promoters and enhancers that control gene expression in specific regions and at specific times, limits usage of these techniques. However, progress in in utero systems of electroporation into mouse embryos has opened a new window, permitting new approaches to answering important questions. Simple injection of plasmid DNA solution and application of electrical current to mouse embryos results in transient area- and time-dependent transfection. Further modification of the technique, arising from variations in types of electrodes used, has made it possible to control the relative size of the region of transfection, which can vary from a few cells to entire tissues. Thus, this technique is a powerful means not only of characterizing gene function in various settings, but also of tracing the migratory routes of cells, due to its high efficiency and the localization of gene expression it yields. We summarize here some of the potential uses and advantages of this technique for developmental neuroscience research.
小鼠基因操作,如基因敲除、基因敲入和转基因小鼠的产生,为分析发育过程中众多基因的功能提供了出色的系统。然而,缺乏在特定区域和特定时间控制基因表达的特异性启动子和增强子,限制了这些技术的应用。不过,小鼠胚胎子宫内电穿孔系统的进展打开了一扇新窗口,允许采用新方法来回答重要问题。将质粒DNA溶液简单注射到小鼠胚胎并施加电流,会导致瞬时的区域和时间依赖性转染。由于所使用电极类型的差异,对该技术的进一步改进使得控制转染区域的相对大小成为可能,转染区域大小可从几个细胞到整个组织不等。因此,由于其高效率以及所产生的基因表达定位,该技术不仅是在各种情况下表征基因功能的有力手段,也是追踪细胞迁移路线的有力手段。我们在此总结该技术在发育神经科学研究中的一些潜在用途和优势。