Properties of an 'LNA'-modified ricin RNA aptamer
Properties of an 'LNA'-modified ricin RNA aptamer
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DOI：
10.1016/j.bbrc.2012.01.127
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发表时间：
2012-03-02
影响因子：
3.1
通讯作者：
Fuerste, Jens P.
中科院分区：
文献类型：
--
作者：
Foerster, Charlotte;Zydek, Martin;Fuerste, Jens P.
"锁核酸"（LNA）是含有2'-O-4'C-亚甲基-β-D-呋喃核糖的糖修饰的核酸。用LNA取代RNA导致增强的热稳定性。适体是通过"SELEX"方法从大文库池中选择用于特异性靶结合的核酸。将修饰的核酸引入适体可以提高其稳定性。蓖麻毒素A链RNA适体的茎区被锁核酸取代。与天然RNA对应物相比，检查LNA取代的适体的不同构建体的热稳定性、结合活性、折叠和RNA酶敏感性。LNA修饰的适体在靶结合中是活性的，而环区域和相邻的茎核苷酸保持未取代。与天然RNA适体相比，LNA取代的适体的热稳定性和RNA酶抗性增强。这项研究支持的方法，取代适体茎区的LNA，并离开环区未修饰，这是负责配体结合。因此，LNA具有开发新的稳定化核酸的令人鼓舞的潜力，并将促进未来的诊断和治疗应用。(C)2012 Elsevier Inc. All rights reserved.
'Locked nucleic acids' (LNAs) are sugar modified nucleic acids containing the 2'-O-4'C-methylene-beta-D-ribofuranoses. The substitution of RNAs with LNAs leads to an enhanced thermostability. Aptamers are nucleic acids, which are selected for specific target binding from a large library pool by the 'SELEX' method. Introduction of modified nucleic acids into aptamers can improve their stability. The stem region of a ricin A chain RNA aptamer was substituted by locked nucleic acids. Different constructs of the LNA-substituted aptamers were examined for their thermostability, binding activity, folding and RNase sensitivity as compared to the natural RNA counterpart. The LNA-modified aptamers were active in target binding, while the loop regions and the adjacent stem nucleotides remained unsubstituted. The thermostability and RNase resistance of LNA substituted aptamers were enhanced as compared to the native RNA aptamer. This study supports the approach to substitute the aptamer stem region by LNAs and to leave the loop region unmodified, which is responsible for ligand binding. Thus, LNAs possess an encouraging potential for the development of new stabilized nucleic acids and will promote future diagnostic and therapeutic applications. (C) 2012 Elsevier Inc. All rights reserved.