Manipulation of MicroRNAs in Cultured Mouse Embryos: Applications for Developmental Toxicology.
Manipulation of MicroRNAs in Cultured Mouse Embryos: Applications for Developmental Toxicology.
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DOI：
10.1007/978-1-4939-7883-0_10
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发表时间：
2018
影响因子：
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通讯作者：
Fuqiang Yuan;Shao-yu Chen
中科院分区：
文献类型：
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作者：
Fuqiang Yuan;Shao-yu Chen
MicroRNA (miRNA) 是一类小的非编码 RNA，可调节几乎所有基因的表达。 miRNA 参与广泛的细胞和生理过程的调节。已发现 miRNA 失调与多种疾病和病症有关，包括致畸。揭示 miRNA 在致畸过程中功能的重要策略之一是操纵其在发育胚胎特定区域的表达水平和模式。显微注射技术与全胚胎培养（WEC）系统的结合为miRNA在致畸中的作用机制研究提供了良好的平台。在这里，我们描述了一种在培养的小鼠胚胎中研究和操作 miRNA 的方法，并提出了全胚胎培养、miRNA 模拟物或抑制剂的显微注射、胚胎中 microRNA 调节功效的验证以及致畸诱导的细胞凋亡的评估的方案。
MicroRNAs (miRNAs) are a class of small noncoding RNAs that modulate the expression of virtually all genes. miRNAs have been implicated in the regulation of a broad range of cellular and physiologic processes. Dysregulation of miRNAs has been found to be associated with a variety of diseases and disorders, including teratogenesis. One of the important strategies to reveal miRNA function in teratogenesis is the manipulation of their expression levels and patterns in specific regions in the developing embryos. The combination of microinjection technology with the whole embryo culture (WEC) system provides an excellent platform for the mechanistic study of the roles of miRNAs in teratogenesis. Here we describe an approach to investigating and manipulating miRNAs in cultured mouse embryos and present protocols for whole embryo culture, microinjection of miRNA mimics or inhibitors, validation of the efficacy of microRNA modulation in embryos, and assessment of the teratogen-induced apoptosis.