Interactions of psoralen-derivatized oligodeoxyribonucleoside methylphosphonates with vesicular stomatitis virus messenger RNA.
Interactions of psoralen-derivatized oligodeoxyribonucleoside methylphosphonates with vesicular stomatitis virus messenger RNA.
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补骨脂素衍生的寡脱氧核糖核苷甲基膦酸酯与水泡性口炎病毒信使 RNA 的相互作用。
DOI：
10.1089/ard.1994.4.223
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发表时间：
1994
期刊：
影响因子：
--
通讯作者：
Miller,PS
中科院分区：
文献类型：
--
作者：
Levis,JT;Miller,PS
寡核苷酸选择性地与其靶标相互作用的能力是设计反义寡核苷酸的一个重要考虑因素。这在反义低聚物的情况下尤其重要，例如补骨脂素衍生的低聚物，它们可以不可逆地结合到它们的靶标上。我们研究了补骨脂素衍生的一系列反义寡核苷酸甲基膦与水疱性口炎病毒(VSV)的mRNAs的相互作用，这些mRNAs具有高度的序列同源性。为了减少mRNA的二级结构，在低离子强度条件下进行了交联反应。12-聚体，其序列与VSV M-mRNA互补，与N、NS和G中发现的序列部分互补，广泛地与N-Message交联。另一方面，其序列与N-或M-mRNA上的结合位点唯一互补的16-MERS在0°~37°C的温度范围内特异而有效地与其目标mRNAs交联。逆转录酶催化的引物延伸试验表明，其中一个N-特异性寡聚体在N-mRNA上的预期位置发生了交联性并估计了交联度。结果表明，如果选择合适的序列，补骨脂素衍生的低聚核苷甲基膦酸盐能够以序列特异性的方式进行交联，以避免与其他mRNA序列的广泛部分互补。
The ability of oligonucleotides to interact selectively with their targets is an important consideration in the design of antisense oligonucleotides. This is especially important in the case of antisense oligomers, such as psoralen-derivatized oligomers, which can irreversibly bind to their targets. We have studied the interactions of a series of psoralen-derivatized antisense oligonucleoside methylphosphonates with the mRNAs of vesicular stomatitis virus (VSV), mRNAs that have a high degree of sequence homology. Cross-linking reactions were carried out under conditions of low ionic strength in order to reduce mRNA secondary structure. A 12-mer, whose sequence was complementary to VSV M -mRNA and partially complementary to sequences found in N, NS, and G mRNA cross-linked extensively to N-message. On the other hand, 16-mers whose sequences were uniquely complementary to binding sites on N- or M-mRNA specifically and efficiently cross-linked to their targeted mRNAs over the temperature range 0° to 37°C. A reverse transcriptase-catalyzed primer extension assay was used to show that one of the N-specific oligomers cross-linked at the expected site on N-mRNA and to estimate the extent of cross-linking. The results demonstrate that psoralen-derivatized oligonucleoside methylphosphonates can cross-link in a sequence-specific manner if the sequences of these oligomers are chosen carefully so as to avoid extensive partial complementarity with other mRNA sequences.