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关于如何运用卵内电穿孔技术分析基因功能的入门指南。

A primer on using in ovo electroporation to analyze gene function.

作者信息

Krull Catherine E

机构信息

Biological Sciences, University of Missouri-Columbia, Columbia, Missouri 65211, USA.

出版信息

Dev Dyn. 2004 Mar;229(3):433-9. doi: 10.1002/dvdy.10473.

Abstract

The chicken embryo has served as a classic model system for developmental studies due to its easy access for surgical manipulations and a wealth of data about chicken embryogenesis. Notably, the mechanisms controlling limb development have been explored best in the chick. Recently, the method of in ovo electroporation has been used successfully to transfect particular cells/tissues during embryonic development, without the production or infectivity associated with retroviruses. With the sequencing of the chicken genome near completion, this approach will provide a powerful opportunity to examine the function of chicken genes and their counterparts in other species. In ovo electroporation has been most effectively used to date for ectopic or overexpression analyses. However, recent studies indicate that this approach can be used successfully for loss-of-function analyses, including protein knockdown experiments with morpholinos and RNAi. Here, I will discuss parameters for using in ovo electroporation successfully to study developmental processes.

摘要

由于鸡胚胎易于进行手术操作且有大量关于鸡胚胎发育的数据,它一直是发育研究的经典模型系统。值得注意的是,在鸡中对控制肢体发育的机制进行了最为深入的探索。最近,胚胎内电穿孔方法已成功用于在胚胎发育过程中转染特定细胞/组织,而不会产生与逆转录病毒相关的产物或感染性。随着鸡基因组测序接近完成,这种方法将为研究鸡基因及其在其他物种中的对应基因的功能提供强大的机会。迄今为止,胚胎内电穿孔最有效地用于异位或过表达分析。然而,最近的研究表明,这种方法可成功用于功能缺失分析,包括使用吗啉代寡核苷酸和RNA干扰进行的蛋白质敲低实验。在此,我将讨论成功使用胚胎内电穿孔研究发育过程的参数。

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