CAMS Key Laboratory of Antiviral Drug Research, Beijing Key Laboratory of Antimicrobial Agents, NHC Key Laboratory of Biotechnology of Antibiotics, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100050, China.
Beijing Key Laboratory of Drug Resistance Tuberculosis Research, Beijing Tuberculosis and Thoracic Tumor Research Institute, and Beijing Chest Hospital, Capital Medical University, Beijing, China.
Talanta. 2024 Jan 15;267:125213. doi: 10.1016/j.talanta.2023.125213. Epub 2023 Sep 16.
The binding site-oriented SELEX strategy is an effective way to promote the functional evolution of aptamers. Here we report a novel aptamer screening strategy (Differentiated-competition Capillary Electrophoresis-SELEX), which enables candidate aptamers to be directionally induced to bind to different active sites of RecA. In this strategy, we introduce two competing binding factors into the "binding - dissociation" dynamic equilibrium system of RecA and ssDNA libraries. Due to the completely different binding mechanism of the competitive factor and the ssDNA library, it exerts different interference on the binding of RecA and the ssDNA library, which directed the binding site of aptamer candidates during the SELEX process. Multifunctional aptamers with high affinity and specificity were found to inhibit RecA activity by binding to different active sites. In conclusion, the SELEX method developed in our current study have identified a variety of biologically functional aptamers with relatively well-defined binding sites that regulate RecA protein activity, which has potential applications and future prospects for accurate screening of biological functional aptamers.
基于结合位的 SELEX 策略是促进适体功能进化的有效方法。在这里，我们报告了一种新的适体筛选策略（差异化竞争毛细管电泳-SELEX），它可以定向诱导候选适体与 RecA 的不同活性位结合。在该策略中，我们将两种竞争结合因子引入 RecA 和 ssDNA 文库的“结合-解离”动态平衡体系中。由于竞争因子和 ssDNA 文库的结合机制完全不同，它对 RecA 和 ssDNA 文库的结合产生了不同的干扰，从而在 SELEX 过程中指导了适体候选物的结合位。通过与不同的活性位结合，我们发现具有高亲和力和特异性的多功能适体能抑制 RecA 活性。总之，我们目前研究中开发的 SELEX 方法已经鉴定出多种具有相对明确结合位的具有生物功能的适体，这些适体可以调节 RecA 蛋白活性，为准确筛选具有生物功能的适体提供了应用前景和未来展望。
