Taniguchi Yosuke, Kurose Yusuke, Nishioka Takamasa, Nagatsugi Fumi, Sasaki Shigeki
Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka 812-8582, Japan.
Nucleic Acids Symp Ser (Oxf). 2009(53):167-8. doi: 10.1093/nass/nrp084.
We have previously reported that the 2-amino-6-vinylpurine (AVP) in oligonucleotide (ODN) showed the highly selective and effective cross-linking reaction toward cytosine within target complementary sequences. Recently, we revealed that the PEGylated ODN containing AVP analogues exhibited the effective antisense effect in the cultured cell. However, subsequent studies showed that the cross-linking ability of AVP for cytosine was affected by the target RNA sequences. We therefore developed new intelligent nucleoside analogues connected AVP to sugar through the ethylene linker (et-AVP), that exhibited highly effective cross-linking ability to rC in the target RNA. In this paper, we described the synthesis of the PEGylated ODN containing cross-linking nucleoside analogues and properties of it about RNase H activity and antisense effect in cell lysate. As a result, the PEGylated ODN containing et-AVP showed the effective antisense effect in the non-cell system for targeting the luciferase mRNA by non-RNase-H mechanism.
我们之前报道过，寡核苷酸（ODN）中的2-氨基-6-乙烯基嘌呤（AVP）对靶互补序列中的胞嘧啶表现出高度选择性和有效的交联反应。最近，我们发现含有AVP类似物的聚乙二醇化ODN在培养细胞中表现出有效的反义效应。然而，随后的研究表明，AVP对胞嘧啶的交联能力受靶RNA序列影响。因此，我们开发了通过乙烯连接子（et-AVP）将AVP与糖连接的新型智能核苷类似物，其对靶RNA中的rC表现出高效交联能力。在本文中，我们描述了含有交联核苷类似物的聚乙二醇化ODN的合成及其在细胞裂解物中的RNase H活性和反义效应特性。结果，含有et-AVP的聚乙二醇化ODN通过非RNase-H机制在非细胞系统中对萤光素酶mRNA靶向表现出有效的反义效应。
