Studying RNA-Binding Protein Interactions with Target mRNAs in Eukaryotic Cells: Native Ribonucleoprotein Immunoprecipitation (RIP) Assays
Studying RNA-Binding Protein Interactions with Target mRNAs in Eukaryotic Cells: Native Ribonucleoprotein Immunoprecipitation (RIP) Assays
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DOI：
10.1007/978-1-4939-2253-6_14
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发表时间：
2015-01-01
期刊：
影响因子：
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通讯作者：
Brody, Jonathan R.
中科院分区：
文献类型：
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作者：
Cozzitorto, Joseph A.;Jimbo, Masaya;Brody, Jonathan R.
mRNA 的转录后调控可以有效地决定真核细胞中的蛋白质表达模式。这种调节模式通过 mRNA 转录物中的顺式作用调节区域发生，介导与反式作用 RNA 结合蛋白 (RBP) 的直接相互作用。这种 mRNA/蛋白质相互作用可以通过多种方式进行研究，从体外到体内，通过信使核糖核蛋白免疫沉淀（mRNP-IP 或 RIP）测定。这种改良的免疫沉淀方法是确定不同细胞应激源下特定 RBP 对基因表达调节的重要且灵敏的方法。
Post-transcriptional regulation of mRNA can potently dictate protein expression patterns in eukaryotic cells. This mode of regulation occurs through cis-acting regulatory regions in the mRNA transcript that mediate direct interactions with trans-acting RNA-binding proteins (RBPs). This mRNA/protein interaction can be studied in numerous ways that range from in vitro to in vivo through messenger ribonucleoprotein immunoprecipitation (mRNP-IP or RIP) assays. This modified immunoprecipitation approach is an important and sensitive method to determine the regulation of gene expression by specific RBPs under different cellular stressors.