Laboratory of Ribosome Structure and Functions, Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Prospect Lavrentieva 8, Novosibirsk 630090, Russia.
Molecules. 2013 Nov 25;18(12):14455-69. doi: 10.3390/molecules181214455.
Introduction of reporter groups at designed RNA sites is a widely accepted approach to gain information about the molecular environment of RNAs in their complexes with other biopolymers formed during various cellular processes. A general approach to obtain RNAs bearing diverse reporter groups at designed locations is based on site-specific insertion of groups containing primary aliphatic amine functions (amino linkers) with their subsequent selective derivatization by appropriate chemicals. This article is a brief review on methods for site-specific introduction of amino linkers in different RNAs. These methods comprise: (i) incorporation of a nucleoside carrying an amino-linker or a function that can be substituted with it into oligoribonucleotides in the course of their chemical synthesis; (ii) assembly of amino linker-containing RNAs from short synthetic fragments via their ligation; (iii) synthesis of amino linker-modified RNAs using T7 RNA polymerase; (iv) insertion of amino linkers into unmodified RNAs at functional groups of a certain type such as the 5'-phosphates and N7 of guanosine residues and (v) introduction of an amino linker into long highly structured RNAs exploiting an approach based on sequence-specific modification of nucleic acids. Particular reporter groups used for derivatization of amino linker-containing RNAs together with types of RNA derivatives obtained and fields of their application are presented.
在设计的 RNA 位点引入报告基团是一种广泛接受的方法，可以获取关于 RNA 与其在各种细胞过程中与其他生物聚合物形成的复合物中分子环境的信息。获得在设计位置带有各种报告基团的 RNA 的一般方法是基于在特定位置插入含有伯脂肪族胺功能的基团（氨基接头），并通过适当的化学物质对其进行选择性衍生化。本文简要综述了在不同 RNA 中特异性引入氨基接头的方法。这些方法包括：（i）在寡核糖核苷酸的化学合成过程中掺入携带氨基接头或可与其取代的核苷；（ii）通过连接将含有氨基接头的 RNA 从短的合成片段组装而成；（iii）使用 T7 RNA 聚合酶合成氨基接头修饰的 RNA；（iv）在特定类型的官能团（如 5' - 磷酸和鸟苷残基的 N7）上将氨基接头插入未修饰的 RNA 中；（v）利用基于核酸序列特异性修饰的方法将氨基接头引入长的高度结构化的 RNA 中。本文介绍了用于衍生含氨基接头的 RNA 的特定报告基团以及获得的 RNA 衍生物的类型及其应用领域。
