INHIBITION OF TRANSLATION INITIATION BY ANTISENSE OLIGONUCLEOTIDES VIA AN RNASE-H INDEPENDENT MECHANISM
INHIBITION OF TRANSLATION INITIATION BY ANTISENSE OLIGONUCLEOTIDES VIA AN RNASE-H INDEPENDENT MECHANISM
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DOI：
10.1093/nar/19.5.1113
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发表时间：
1991-03-11
影响因子：
14.9
通讯作者：
TOULME, JJ
中科院分区：
文献类型：
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作者：
BOIZIAU, C;KURFURST, R;TOULME, JJ
我们已经使用α-寡聚体作为互补于兔β-珠蛋白mRNA的三个不同序列的反义寡核苷酸：邻近帽位点的区域、跨越AUG起始密码子的区域或编码区中的序列。 这些α-寡核苷酸是用游离的5 ′ OH基团或连接到吖啶衍生物上合成的。 在无细胞提取物和非洲爪蟾卵母细胞中研究了这些寡核苷酸对mRNA翻译的影响。 在兔网织红细胞裂解物和小麦胚芽提取物中，靶向帽位点和起始密码子的低聚物以剂量依赖性方式减少β-珠蛋白合成，而靶mRNA保持完整。 在兔网织红细胞裂解物中，抗cap α-寡聚体甚至比其β-对应物更有效。 与此相反，只有α-寡聚体，连接到吖啶衍生物，互补的帽区显示显着的反义特性在非洲爪蟾卵母细胞。 因此，翻译的起始可以被不是RNase-H底物的寡核苷酸/RNA杂合体阻止。
We have used alpha-oligomers as antisense oligonucleotides complementary to three different sequences of the rabbit beta-globin mRNA: a region adjacent to the cap site, a region spanning the AUG initiation codon or a sequence in the coding region. These alpha-oligonucleotides were synthesized either with a free 5' OH group or linked to an acridine derivative. The effect of these oligonucleotides on mRNA translation was investigated in cell-free extracts and in Xenopus oocytes. In rabbit reticulocyte lysate and in wheat germ extracts oligomers targeted to the cap site and the initiation codon reduced beta-globin synthesis in a dose-dependent manner, whereas the target mRNA remained intact. The anti-cap alpha-oligomer was even more efficient that its beta-counterpart in rabbit reticulocyte lysate. In contrast, only the alpha-oligomer, linked to the acridine derivative, complementary to the cap region displayed significant antisense properties in Xenopus oocytes. Therefore initiation of translation can be arrested by oligonucleotide/RNA hybrids which are not substrates for RNase-H.