Synthesis and hybridization properties of oligonucleotide perylene conjugates: influence of the conjugation parameters on triplex and duplex stabilities
Synthesis and hybridization properties of oligonucleotide perylene conjugates: influence of the conjugation parameters on triplex and duplex stabilities
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DOI：
10.1039/b410695d
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发表时间：
2004-01-01
影响因子：
3.2
通讯作者：
Asseline, U
中科院分区：
文献类型：
--
作者：
Aubert, Y;Asseline, U
本文报道了一种2‘-脱氧寡核苷酸(ODN)的合成。在2‘-脱氧核糖残基的异构体位置上直接或通过丙基连接物的引入，诱导形成两个异构体。这些糖衍生物单元的每个纯异构体都在嘧啶类十五聚体的5‘端或内部位置进行了单结合。用吸收光谱研究了这些修饰的ODN与其单链和双链DNA靶标的结合特性。在ODN链的5‘端和内部位置(或同时在5’端和3‘端)进行了在稳定三链和双链结构(涉及丙基连接物的β-异构体)方面最有效的糖二茂单元的双掺入。已经与包括一个或两个通过较长的聚亚甲基链连接到5‘-或3’-(或5‘-和3’-)末端磷酸基上的一个或两个perenes的ODN perene偶联物进行了比较。在稳定三链体和双链体结构方面，含有两个perenes的ODN比只含有一个perenes的ODN更有效，其中，含有两个糖基末端的ODN perene结合物是最有效的。荧光研究的结果表明，当涉及两个perenes的ODN与单链或双链靶标杂交时，荧光信号的强度显著增加。这种荧光信号强度的增加可以作为杂交的证据。
We report here the synthesis of oligo-2'-deoxyribonucleotides (ODNs) conjugated with perylene. Introduction of perylene, coupled either directly or via a propyl linker to the anomeric position of a 2'-deoxyribose residue, induces the formation of two anomers. Single incorporations of each pure anomer of these sugar perylene units have been performed at either the 5'-end or an internal position of a pyrimidic pentadecamer. The binding properties of these modified ODNs with their single- and double-stranded DNA targets were studied by absorption spectroscopy. Double incorporations of the sugar perylene unit most efficient at stabilizing the triplex and duplex structures (the beta-anomer involving the propyl linker) have been performed at both the 5'-end and at an internal position (or both the 5'- and 3'-ends) of the ODN chain. Comparison has been made with ODN perylene conjugates involving either one or two perylenes attached via a longer polymethylene chain to either the 5'- or 3'- (or both the 5'- and 3'-) terminal phosphate groups. The ODNs involving two perylenes are more efficient at stabilizing the triplex and the duplex structures than the ODNs involving only one perylene and, among these, the ODN perylene conjugate involving two sugar perylene units attached at both termini is the most efficient. The results of the fluorescence studies have shown an important increase in the intensity of the fluorescent signal upon hybridization of the ODNs involving two perylenes with either the single- or the double-stranded targets. This increase in the intensity of the fluorescent signal could be used as proof of the hybridization.