Mereau Agnès, Hardy Serge
UMR 6061 CNRS-Université de Rennes 1, Rennes, France.
Methods Mol Biol. 2012;917:347-68. doi: 10.1007/978-1-61779-992-1_20.
Alternative splicing, the process by which distinct mature mRNAs can be produced from a single primary transcript, is a key mechanism to increase the organism complexity. The generation of alternative splicing pattern is a means to expand the proteome diversity and also to control gene expression through the regulation of mRNA abundance. Alternative splicing is therefore particularly prevalent during development and accordingly numerous splicing events are regulated in a tissue or temporal manner. To study the roles of alternative splicing during developmental processes and decipher the molecular mechanisms that underlie temporal and spatial regulation, it is important to develop in vivo whole animal studies. In this chapter, we present the advantages of using the amphibian Xenopus as a fully in vivo model to study alternative splicing and we describe the experimental procedures that can be used with Xenopus laevis embryos and oocytes to define the cis-regulatory elements and identify the associated trans-acting factors.
可变剪接是指从单个初级转录本产生不同成熟mRNA的过程，是增加生物体复杂性的关键机制。可变剪接模式的产生是扩展蛋白质组多样性以及通过调节mRNA丰度来控制基因表达的一种方式。因此，可变剪接在发育过程中尤为普遍，相应地，许多剪接事件以组织或时间方式受到调控。为了研究可变剪接在发育过程中的作用并破译时空调控背后的分子机制，开展体内全动物研究很重要。在本章中，我们阐述了使用两栖动物非洲爪蟾作为完全体内模型来研究可变剪接的优势，并描述了可用于非洲爪蟾胚胎和卵母细胞以定义顺式调控元件并鉴定相关反式作用因子的实验程序。
