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用于SELEX适体筛选的寡核苷酸文库中固定引物区域的新设计。

A New Design for the Fixed Primer Regions in an Oligonucleotide Library for SELEX Aptamer Screening.

作者信息

Wang Bin

机构信息

Department of Chemistry, Marshall University, Huntington, WV, United States.

出版信息

Front Chem. 2020 Jun 5;8:475. doi: 10.3389/fchem.2020.00475. eCollection 2020.

Abstract

Single-stranded DNA or RNA oligonucleotides, often called aptamers, can bind to a molecular target with both high affinity and selectivity due to their distinct three-dimensional structures. A technique called systematic evolution of ligands by exponential enrichment (SELEX) is used to screen aptamers from a random DNA or RNA pool, or library. The traditionally-designed oligonucleotides in libraries contain a randomized central region along with a fixed primer region at each end for amplifying target-bound central sequences. The single-stranded forward and reverse primer sequences may interfere with target-binding to the central region, resulting in a partial or complete loss of high-affinity aptamers during the SELEX process. To address this issue, researchers have modified the traditional oligonucleotide libraries and developed new types of oligonucleotide libraries; however, these approaches come with various limitations. The author proposes a new design that uses a conformation-changeable sequence as primers, which may open a new avenue for developing an optimized aptamer sequence with both high affinity for, and selective binding to, a particular target via SELEX.

摘要

单链DNA或RNA寡核苷酸,通常称为适配体,由于其独特的三维结构,能够以高亲和力和选择性与分子靶标结合。一种称为指数富集配体系统进化(SELEX)的技术用于从随机DNA或RNA文库中筛选适配体。文库中传统设计的寡核苷酸包含一个随机化的中心区域以及两端用于扩增与靶标结合的中心序列的固定引物区域。单链正向和反向引物序列可能会干扰靶标与中心区域的结合,导致在SELEX过程中高亲和力适配体部分或完全丢失。为了解决这个问题,研究人员对传统寡核苷酸文库进行了修改并开发了新型寡核苷酸文库;然而,这些方法存在各种局限性。作者提出了一种新的设计,使用构象可变序列作为引物,这可能为通过SELEX开发对特定靶标具有高亲和力和选择性结合的优化适配体序列开辟一条新途径。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6492/7291870/0954028c4019/fchem-08-00475-g0001.jpg

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