Efficient reverse transcription using locked nucleic acid nucleotides towards the evolution of nuclease resistant RNA aptamers.
Efficient reverse transcription using locked nucleic acid nucleotides towards the evolution of nuclease resistant RNA aptamers.
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DOI：
10.1371/journal.pone.0035990
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Veedu RN
中科院分区：
文献类型：
--
作者：
Crouzier L;Dubois C;Edwards SL;Lauridsen LH;Wengel J;Veedu RN
修饰的核苷酸越来越多地用于适配体的从头选择，以增强其药物样特征并消除对耗时的基于试错法的选择后修饰的需要。锁核酸（Locked nucleic acid，LNA）是一种重要的核酸类似物，因其独特的性质而被广泛应用于治疗性寡核苷酸的构建。LNA修饰的RNA适体的进化需要一种有效的逆转录方法来PCR富集所选的RNA适体候选物。建立这一关键步骤是进行LNA修饰的RNA适体选择的先决条件。在这项研究中，三种不同的逆转录酶进行了研究对LNA核苷酸的酶识别。研究LNA核苷酸的掺入以及阅读能力以充分理解酶识别的局限性。我们发现SuperScript® III逆转录酶是一种有效识别LNA核苷酸的酶，使其成为用于从头选择含有RNA适体的LNA的主要候选物。
Modified nucleotides are increasingly being utilized in the de novo selection of aptamers for enhancing their drug-like character and abolishing the need for time consuming trial-and-error based post-selection modifications. Locked nucleic acid (LNA) is one of the most prominent and successful nucleic acid analogues because of its remarkable properties, and widely explored as building blocks in therapeutic oligonucleotides. Evolution of LNA-modified RNA aptamers requires an efficient reverse transcription method for PCR enrichment of the selected RNA aptamer candidates. Establishing this key step is a pre-requisite for performing LNA-modified RNA aptamer selection. In this study three different reverse transcriptases were investigated towards the enzymatic recognition of LNA nucleotides. Both incorporation as well as reading capabilities of the LNA nucleotides was investigated to fully understand the limitations of the enzymatic recognition. We found that SuperScript® III Reverse Transcriptase is an efficient enzyme for the recognition of LNA nucleotides, making it a prime candidate to be used in de novo selection of LNA containing RNA aptamers
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影响因子：
2.1
作者：
Koshkin, AA;Singh, SK;Wengel, J
通讯作者：
Wengel, J
影响因子：
3.2
作者：
Veedu, Rakesh N.;Vester, Birte;Wengel, Jesper
通讯作者：
Wengel, Jesper
影响因子：
3.2
作者：
Veedu, Rakesh N.;Vester, Birte;Wengel, Jesper
通讯作者：
Wengel, Jesper
影响因子：
1.8
作者：
Obika, S;Nanbu, D;Imanishi, T
通讯作者：
Imanishi, T
影响因子：
4.8
作者：
Ruckman, J;Green, LS;Janjic, N
通讯作者：
Janjic, N