Key Laboratory of Specialty Agri-Products Quality and Hazard Controlling Technology of Zhejiang Province, College of Life Sciences, China Jiliang University, Hangzhou 310018, China.
Shenzhen Key Laboratory of Advanced Thin Films and Applications, College of Physics and Optoelectronic Engineering, Shenzhen University, Shenzhen 518060, China.
Biosensors (Basel). 2024 Jul 18;14(7):350. doi: 10.3390/bios14070350.
Aptamers are nucleic acid sequences that specifically bind with target molecules and are vital to applications such as biosensing, drug development, disease diagnostics, etc. The traditional selection procedure of aptamers is based on the Systematic Evolution of Ligands by an Exponential Enrichment (SELEX) process, which relies on repeating cycles of screening and amplification. With the rapid development of aptamer applications, RNA and XNA aptamers draw more attention than before. But their selection is troublesome due to the necessary reverse transcription and transcription process (RNA) or low efficiency and accuracy of enzymes for amplification (XNA). In light of this, we review the recent advances in aptamer selection methods and give an outlook on future development in a non-SELEX approach, which simplifies the procedure and reduces the experimental costs. We first provide an overview of the traditional SELEX methods mostly designed for screening DNA aptamers to introduce the common tools and methods. Then a section on the current screening methods for RNA and XNA is prepared to demonstrate the efforts put into screening these aptamers and the current difficulties. We further predict that the future trend of aptamer selection lies in non-SELEX methods that do not require nucleic acid amplification. We divide non-SELEX methods into an immobilized format and non-immobilized format and discuss how high-resolution partitioning methods could facilitate the further improvement of selection efficiency and accuracy.
适体是一种能够特异性结合靶分子的核酸序列，在生物传感、药物开发、疾病诊断等应用中具有重要作用。适体的传统筛选程序是基于指数富集的配体系统进化（SELEX）过程，该过程依赖于筛选和扩增的重复循环。随着适体应用的快速发展，RNA 和 XNA 适体比以前受到了更多的关注。但由于其必要的反转录和转录过程（RNA）或扩增酶的低效率和准确性（XNA），它们的选择比较麻烦。有鉴于此，我们综述了适体选择方法的最新进展，并对非 SELEX 方法的未来发展进行了展望，该方法简化了程序并降低了实验成本。我们首先概述了主要用于筛选 DNA 适体的传统 SELEX 方法，以介绍常用的工具和方法。然后准备了一个关于 RNA 和 XNA 当前筛选方法的部分，以展示筛选这些适体的努力和当前的困难。我们进一步预测，适体选择的未来趋势在于不需要核酸扩增的非 SELEX 方法。我们将非 SELEX 方法分为固定化格式和非固定化格式，并讨论了如何使用高分辨率分离方法来提高选择效率和准确性。
