The shorter the better: reducing fixed primer regions of oligonucleotide libraries for aptamer selection.
The shorter the better: reducing fixed primer regions of oligonucleotide libraries for aptamer selection.
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DOI：
10.3390/molecules14041353
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发表时间：
2009-03-27
期刊：
影响因子：
--
通讯作者：
Clawson GA
中科院分区：
文献类型：
--
作者：
Pan W;Clawson GA
寡核苷酸适体是高度结构化的DNA或RNA分子或其修饰形式，其可以以与抗体相当的特异性亲和力结合靶标。它们通过称为SELEX（指数富集配体系统进化）的体外选择过程进行鉴定，以识别从小分子到蛋白质以及从培养细胞到整个生物体的各种靶标。适体具有许多期望的性质，例如易于合成、稳定性、稳健性和缺乏免疫原性。标准的SELEX文库需要两个引物，一个在中心随机结构域的每一侧，以通过PCR或RT-PCR扩增靶结合序列。然而，这些引物序列因其性质而引起非特异性结合，并且已报道导致大量假阳性结合序列，或干扰随机区域内序列的结合。本文综述了在SELEX过程中消除固定引物干扰的方法。
Oligonucleotide aptamers are highly structured DNA or RNA molecules, or modified versions thereof, that can bind to targets with specific affinities comparable to antibodies. They are identified through an in vitro selection process termed SELEX (Systematic Evolution of Ligands by EXponential enrichment) to recognize a wide variety of targets, from small molecules to proteins, and from cultured cells to whole organisms. Aptamers possess a number of desirable properties, such as ease of synthesis, stability, robustness, and lack of immunogenicity. Standard SELEX libraries require two primers, one on each side of a central random domain, to amplify the target-bound sequences via PCR or RT-PCR. However, these primer sequences cause non-specific binding by their nature, and have been reported to lead to large numbers of false-positive binding sequences, or to interfere with binding of sequences within the random regions. This review is focused on methods which have been developed to eliminate fixed primer interference during the SELEX process.
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