Meikar Oliver, Kotaja Noora
Department of Physiology, Institute of Biomedicine, University of Turku, Kiinamyllynkatu 10, 20520, Turku, Finland.
Methods Mol Biol. 2014;1173:11-25. doi: 10.1007/978-1-4939-0931-5_2.
RNA-protein (RNP) complexes and granules are powerful composites of merged functions and unique properties. The importance of RNPs in carrying out complex tasks in RNA processing and regulation is being increasingly revealed. One of the biggest RNP granules is the chromatoid body (CB) that is believed to orchestrate the RNA posttranscriptional regulation in haploid male germ cells. Here, we describe the CB isolation procedure, from mouse testis. After cross-linking and lysing the cells, the CBs are enriched by slow-speed centrifugation and immunoprecipitated using anti-MVH/DDX4 antibody. The method yields pure fractions of CBs, and it is robust, reproducible and does not require special equipment or abundant starting material. The CB is packed with large amounts of RNA, especially small RNAs. Isolation of the CBs provides a tool to enrich these RNA species.
RNA-蛋白质（RNP）复合物和颗粒是功能融合且具有独特性质的强大复合体。RNP在RNA加工和调控中执行复杂任务的重要性正日益显现。最大的RNP颗粒之一是拟染色体（CB），据信它在单倍体雄性生殖细胞中协调RNA的转录后调控。在此，我们描述从小鼠睾丸中分离CB的程序。在对细胞进行交联和裂解后，通过低速离心富集CB，并使用抗MVH/DDX4抗体进行免疫沉淀。该方法可产生纯净的CB组分，且稳健、可重复，不需要特殊设备或大量起始材料。CB中富含大量RNA，尤其是小RNA。CB的分离提供了一种富集这些RNA种类的工具。
