Department of Chemistry, Texas A&M University, College Station, 77843 TX, USA.
Current address: Department of Chemistry and Biochemistry, Florida State University, 32304, Tallahassee, FL, USA.
Chembiochem. 2022 Dec 16;23(24):e202200520. doi: 10.1002/cbic.202200520. Epub 2022 Nov 22.
Aptamers composed of mirror-image L-(deoxy)ribose nucleic acids, referred to as L-aptamers, are a promising class of RNA-binding reagents. Yet, the selectivity of cross-chiral interactions between L-aptamers and their RNA targets remain poorly characterized, limiting the potential utility of this approach for applications in biological systems. Herein, we carried out the first comprehensive analysis of cross-chiral L-aptamer selectivity using a newly developed "inverse" in vitro selection approach that exploits the genetic nature of the D-RNA ligand. By employing a library of more than a million target-derived sequences, we determined the RNA sequence and structural preference of a model L-aptamer and revealed previously unidentified and potentially broad off-target RNA binding behaviors. These results provide valuable information for assessing the likelihood and consequences of potential off-target interactions and reveal strategies to mitigate these effects. Thus, inverse in vitro selection provides several opportunities to advance L-aptamer technology.
由镜像 L-（脱氧）核糖核酸组成的适体，称为 L-适体，是一类很有前途的 RNA 结合试剂。然而，L-适体与其 RNA 靶标之间的交叉手性相互作用的选择性仍未得到很好的描述，限制了该方法在生物系统中的应用潜力。在此，我们利用新开发的“反向”体外选择方法，对 L-适体的交叉手性选择性进行了首次全面分析，该方法利用了 D-RNA 配体的遗传性质。通过使用一个包含超过 100 万个靶序列的文库，我们确定了一个模型 L-适体的 RNA 序列和结构偏好，并揭示了以前未识别的、可能广泛存在的潜在脱靶 RNA 结合行为。这些结果为评估潜在脱靶相互作用的可能性和后果提供了有价值的信息，并揭示了减轻这些影响的策略。因此，反向体外选择为 L-适体技术的发展提供了多个机会。
