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2',5'-寡腺苷酸:反义嵌合体——合成与性质

2',5'-Oligoadenylate:antisense chimeras--synthesis and properties.

作者信息

Lesiak K, Khamnei S, Torrence P F

机构信息

Section on Biomedical Chemistry, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892.

出版信息

Bioconjug Chem. 1993 Nov-Dec;4(6):467-72. doi: 10.1021/bc00024a008.

Abstract

We have synthesized a novel bioconjugate which joins an antisense oligonucleotide to a unique and potent inhibitor of translation,pn5'A2'(p5'A2')mp5'A(2-5A). Two residues of 4-hydroxybutyl phosphate were employed as linkers to attach the 2',5'-oligoadenylate moiety through its 2'-terminus to the 5'-terminus of the chosen antisense sequence, (dT)20. The syntheses were carried on a solid support according to the phosphite triester method of DNA synthesis (Letsinger, R.L., Lunsford, W.B. (1976) J. Am. Chem. Soc. 98, 3655-3661; Beaucage, S.L., and Caruthers, M.H. (1981) Tetrahedron Lett. 22, 1859-1862). The generated 2-5A antisense chimeras retained both the ability of the 2-5A molecule to activate the 2-5A-dependent RNase as well as the ability of the oligo(dT) moiety to hybridize to the complementary poly(A). Moreover, the chimera, when annealed to its target nucleic acid sequence, was still effectively bound to the 2-5A-dependent nuclease. The methodology described represents a new approach to the selective modulation of mRNA expression.

摘要

我们合成了一种新型生物共轭物,它将反义寡核苷酸与一种独特且有效的翻译抑制剂pn5'A2'(p5'A2')mp5'A(2-5A)连接起来。使用两个4-羟基丁基磷酸残基作为连接子,通过其2'-末端将2',5'-寡腺苷酸部分连接到所选反义序列(dT)20的5'-末端。合成是根据DNA合成的亚磷酸三酯法在固相载体上进行的(莱辛格,R.L.,伦斯福德,W.B.(1976年)《美国化学会志》98,3655 - 3661;博卡奇,S.L.,和卡拉瑟斯,M.H.(1981年)《四面体快报》22,1859 - 1862)。所生成的2-5A反义嵌合体既保留了2-5A分子激活2-5A依赖性核糖核酸酶的能力,也保留了寡聚(dT)部分与互补聚(A)杂交的能力。此外,该嵌合体与靶核酸序列退火后,仍能有效地与2-5A依赖性核酸酶结合。所描述的方法代表了一种选择性调节mRNA表达的新方法。

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