一种使用FMTRIP和邻近连接原位定量RNA-蛋白质相互作用的新方法。
A Novel Method to Quantify RNA-Protein Interactions In Situ Using FMTRIP and Proximity Ligation.
作者信息
Zurla C, Jung J, Blanchard E L, Santangelo P J
机构信息
Wallace H Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, 313 Ferst Drive, UA Whitaker Blgd, Atlanta, GA, 30332, USA.
出版信息
Methods Mol Biol. 2017;1468:155-70. doi: 10.1007/978-1-4939-4035-6_12.
RNA binding proteins (RBP) and small RNAs regulate the editing, localization, stabilization, translation, and degradation of ribonucleic acids (RNAs) through their interactions with specific cis-acting elements within target RNAs. Here, we describe a novel method to detect protein-mRNA interactions, which combines FLAG-peptide modified, multiply-labeled tetravalent RNA imaging probes (FMTRIPs) with proximity ligation (PLA), and rolling circle amplification (RCA). This assay detects native RNA in a sequence specific and single RNA sensitive manner, and PLA allows for the quantification and localization of protein-mRNA interactions with single-interaction sensitivity.
RNA结合蛋白（RBP）和小RNA通过与靶RNA内特定的顺式作用元件相互作用，调节核糖核酸（RNA）的编辑、定位、稳定性、翻译和降解。在此，我们描述了一种检测蛋白质与mRNA相互作用的新方法，该方法将FLAG肽修饰的、多重标记的四价RNA成像探针（FMTRIPs）与邻近连接（PLA）和滚环扩增（RCA）相结合。该检测方法以序列特异性和单RNA敏感性的方式检测天然RNA，并且PLA能够以单相互作用敏感性对蛋白质与mRNA的相互作用进行定量和定位。
Zotero 插件