Barckmann Bridlin, Dufourt Jérémy, Simonelig Martine
mRNA Regulation and Development, Institute of Human Genetics, UMR9002 CNRS-Université de Montpellier, Montpellier Cedex 5, France.
Methods Mol Biol. 2018;1720:89-110. doi: 10.1007/978-1-4939-7540-2_7.
Piwi-interacting RNAs (piRNAs) are a class of small noncoding RNAs bound to specific Argonaute proteins, the PIWI proteins. piRNAs target mRNAs by complementarity to silence them; they play an important role in the repression of transposable elements in the germ line of many species. piRNAs and PIWI proteins are also involved in diverse biological processes through their role in the regulation of cellular mRNAs. In the Drosophila embryo, they contribute to the maternal mRNA decay occurring during the maternal-to-zygotic transition. CLIP (UV cross-linking and immunoprecipitation) techniques have been used to identify target mRNAs of Argonaute proteins. Here we describe the iCLIP (individual-nucleotide resolution CLIP) protocol that we have adapted for the PIWI protein Aubergine in Drosophila embryos.
Piwi相互作用RNA（piRNA）是一类与特定AGO蛋白（PIWI蛋白）结合的小非编码RNA。piRNA通过互补作用靶向mRNA以使其沉默；它们在许多物种生殖系中对转座元件的抑制中发挥重要作用。piRNA和PIWI蛋白还通过其在细胞mRNA调控中的作用参与多种生物学过程。在果蝇胚胎中，它们有助于母源-合子转变期间发生的母源mRNA降解。CLIP（紫外线交联和免疫沉淀）技术已用于鉴定AGO蛋白的靶mRNA。在此，我们描述了我们已适用于果蝇胚胎中PIWI蛋白茄子的iCLIP（单核苷酸分辨率CLIP）方案。
