A new usage of functionalized oligodeoxynucleotide probe for site-specific modification of a guanine base within RNA.
A new usage of functionalized oligodeoxynucleotide probe for site-specific modification of a guanine base within RNA.
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DOI：
10.1093/nar/gkp930
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发表时间：
2010-03
影响因子：
14.9
通讯作者：
Sasaki S
中科院分区：
文献类型：
--
作者：
Onizuka K;Taniguchi Y;Sasaki S
RNA的定点修饰对于研究RNA的结构、功能和动力学具有重要意义。最近，我们报道了一种基于寡核苷酸探针中6-硫代鸟苷碱的1-苯基-2-亚甲基-1，3-二酮单元的官能团转移反应对RNA进行序列和胞嘧啶选择性化学修饰的新方法。在本研究中，我们描述了当反应在碱性条件下进行时，官能团转移速率大大提高，选择性转移到鸟嘌呤碱上。详细研究表明，Rg的2-氨基是官能团转移反应的反应物。作为这种有效的官能团转移反应的潜在应用，以较高的鸟嘌呤和位置选择性成功地将一个相对较大的荧光基团转移到了RNA的目标鸟嘌呤碱基上。这种高效、高选择性的官能团转移反应将为RNA的研究提供新的契机。
Site-specific modification of RNA is of great significance to investigate RNA structure, function and dynamics. Recently, we reported a new method for sequence- and cytosine-selective chemical modification of RNA based on the functional group transfer reaction of the 1-phenyl-2-methylydene-1,3-diketone unit of the 6-thioguanosine base incorporated in the oligodeoxynucleotide probe. In this study, we describe that the functionality transfer rate is greatly enhanced and the selectivity is shifted to the guanine base when the reaction is performed under alkaline conditions. Detailed investigation indicated that the 2-amino group of the enolate form of rG is the reactant of the functionality transfer reaction. As a potential application of this efficient functionality transfer reaction, a pyrene group as a relatively large fluorescent group was successfully transferred to the target guanine base of RNA with a high guanine and site selectivity. This functionality transfer reaction with high efficiency and high site-selectivity would provide a new opportunity as a unique tool for the study of RNA.
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