Sequence-specific targeting of RNA with an oligonucleotide-neomycin conjugate
Sequence-specific targeting of RNA with an oligonucleotide-neomycin conjugate
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DOI：
10.1021/bc060249r
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发表时间：
2007-01-01
影响因子：
4.7
通讯作者：
Arya, Dev P.
中科院分区：
文献类型：
--
作者：
Charles, Irudayasamy;Xi, Hongjuan;Arya, Dev P.
本文报道了在2 '-脱氧尿苷C5位共价连接新霉素的合成方法。概述的合成允许氨基糖苷（新霉素）在存在胸苷（或尿苷）的寡核苷酸（ODN）中的任何给定位点处掺入。将该修饰的碱基并入与七个碱基长的α-八叠球菌素环RNA序列互补的寡核苷酸中，导致增强的双链体杂交。该双链体的T-m增加（Δ T-m = 6 ℃）表明新霉素在双链体沟内的有利相互作用。CD光谱表明，修饰的双链体采用A型确认。ITC测量表明ODN和新霉素与RNA靶标结合的累加效应（K-a = 4.5 × 10（7）M-1）。来自该新霉素-ODN缀合物的杂合双链体的增强的稳定性主要来源于新霉素{Δ Δ H-obs = -7.21 kcal/mol（Δ H-新霉素缀合物-Δ H（非缀合物））}与杂合双链体结合的热贡献。短接头长度允许杂交双链体相对于杂交三链体的选择性稳定。这里描述的结果开辟了新的途径，在核-氨基糖苷-缀合物（N-Ag-C）的设计和合成，其中包括任何数量的氨基糖苷（新霉素）分子每个寡核苷酸可以完成。
The synthesis of neomycin covalently attached at the C5-position of 2'-deoxyuridine is reported. The synthesis outlined allows for incorporation of an aminoglycoside (neomycin) at any given site in an oligonucleotide (ODN) where a thymidine (or uridine) is present. Incorporation of this modified base into an oligonucleotide, which is complementary to a seven-bases-long alpha-sarcin loop RNA sequence, leads to enhanced duplex hybridization. The increase in T-m for this duplex (Delta T-m = 6 degrees C) suggests a favorable interaction of neomycin within the duplex groove. CD spectroscopy shows that the modified duplex adopts an A-type confirmation. ITC measurements indicate the additive effects of ODN and neomycin binding to the RNA target (K-a = 4.5 x 10(7) M-1). The enhanced stability of the hybrid duplex from this neomycin-ODN conjugate originates primarily from the enthalpic contribution of neomycin {Delta Delta H-obs = -7.21 kcal/mol (Delta H-neomycin conjugated - Delta H (nonconjugated))} binding to the hybrid duplex. The short linker length allows for selective stabilization of the hybrid duplex over the hybrid triplex. The results described here open up new avenues in the design and synthesis of nucleo-aminoglycoside-conjugates (N-Ag-C) where the inclusion of any number of aminoglycoside (neomycin) molecules per oligonucleotide can be accomplished.