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卵母细胞特异性敲除:一种研究卵泡发生、卵母细胞成熟、受精和胚胎发生过程中基因功能的新型体内方法。

Oocyte-specific knockout: a novel in vivo approach for studying gene functions during folliculogenesis, oocyte maturation, fertilization, and embryogenesis.

作者信息

Sun Qing-Yuan, Liu Kui, Kikuchi Kazuhiro

机构信息

State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080, China.

出版信息

Biol Reprod. 2008 Dec;79(6):1014-20. doi: 10.1095/biolreprod.108.070409. Epub 2008 Aug 27.

Abstract

Knockout mice have been highly useful tools in helping to understand the functional roles of specific genes in development and diseases. However, in many cases, knockout mice are embryonic lethal, which prevents investigation into a number of important questions, or they display developmental abnormalities, including fertility defects. In contrast, conditional knockout, which is achieved by the Cre-LoxP system, can be used to delete a gene in a specific organ or tissue, or at a specific developmental stage. This technique has advantages over conventional knockout, especially when conventional knockout causes embryonic lethality or when the function of maternal transcripts in early development needs to be defined. Recently, a widely used practice has been used to specifically delete genes of interest in oocytes: Zp3-Cre or Gdf9-Cre transgenic mouse lines, in which Cre-recombinase expression is driven by oocyte-specific zona pellucida 3 (Zp3) promoter or growth differentiation factor 9 (Gdf9) promoter, are crossed with mice bearing floxed target genes. This novel in vivo approach has helped to increase the understanding of the functions of specific genes in folliculogenesis/oogenesis, oocyte maturation, fertilization, and embryogenesis. In this minireview we discuss recent advances in understanding the molecular mechanisms regulating major reproductive and developmental events as revealed by oocyte-specific conditional knockout and perspectives on this technology and related studies.

摘要

基因敲除小鼠在帮助理解特定基因在发育和疾病中的功能作用方面一直是非常有用的工具。然而,在许多情况下,基因敲除小鼠是胚胎致死的,这阻碍了对一些重要问题的研究,或者它们表现出发育异常,包括生育缺陷。相比之下,通过Cre-LoxP系统实现的条件性基因敲除可用于在特定器官或组织中,或在特定发育阶段删除基因。这项技术比传统的基因敲除具有优势,特别是当传统基因敲除导致胚胎致死,或者需要确定早期发育中母体转录本的功能时。最近,一种广泛使用的方法被用于在卵母细胞中特异性删除感兴趣的基因:Zp3-Cre或Gdf9-Cre转基因小鼠品系,其中Cre重组酶的表达由卵母细胞特异性透明带3(Zp3)启动子或生长分化因子9(Gdf9)启动子驱动,与携带floxed靶基因的小鼠杂交。这种新的体内方法有助于增进对特定基因在卵泡发生/卵子发生、卵母细胞成熟、受精和胚胎发生中的功能的理解。在这篇综述中,我们讨论了通过卵母细胞特异性条件性基因敲除揭示的在理解调节主要生殖和发育事件的分子机制方面的最新进展,以及对该技术和相关研究的展望。

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