Analysis of lncRNA-Protein Interactions by RNA-Protein Pull-Down Assays and RNA Immunoprecipitation (RIP)
Analysis of lncRNA-Protein Interactions by RNA-Protein Pull-Down Assays and RNA Immunoprecipitation (RIP)
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DOI：
10.1007/978-1-4939-7371-2_17
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发表时间：
2018-01-01
期刊：
影响因子：
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通讯作者：
Bierhoff, Holger
中科院分区：
文献类型：
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作者：
Bierhoff, Holger
长链非编码RNA（lncRNA）通过将染色质修饰酶靶向不同的基因组位点而在染色质的形成中发挥重要作用。本节涵盖了两种分析lncRNA-蛋白质相互作用的方法。RNA-蛋白质下拉测定使用珠结合蛋白质来捕获体外转录物，或使用固定化合成RNA来结合来自细胞裂解物的蛋白质。在RNA免疫沉淀（RIP）测定中，内源性RNA与感兴趣的蛋白质共免疫沉淀。这两种方法都可以应用于来自增殖和静止细胞的材料，从而提供了对lncRNA-蛋白质相互作用如何在这两种细胞状态之间改变的见解。
Long noncoding RNAs (lncRNAs) have important roles in shaping chromatin by targeting chromatin-modifying enzymes to distinct genomic sites. This section covers two methods to analyze lncRNA-protein interactions. The RNA-protein pull-down assays use either bead-bound proteins to capture in vitro transcripts, or immobilized synthetic RNAs to bind proteins from cell lysates. In the RNA immunoprecipitation (RIP) assay, endogenous RNAs are co-immunoprecipitated with a protein of interest. Both the methods can be applied to material from proliferating and quiescent cells, thus providing insights into how lncRNA-protein interactions are altered between these two cellular states.