Inverse In Vitro Selection Enables Comprehensive Analysis of Cross-Chiral L-Aptamer Interactions.
Inverse In Vitro Selection Enables Comprehensive Analysis of Cross-Chiral L-Aptamer Interactions.
复制标题
反向体外选择可实现跨手性 L 适体相互作用的综合分析。
DOI：
10.1002/cbic.202200520
复制
发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Sczepanski,JonathanT
中科院分区：
文献类型：
--
作者：
Piwko,AlexanderT;Han,Xuan;Kabza,AdamM;Dey,Sougata;Sczepanski,JonathanT
由镜像L-（脱氧）核糖核酸组成的适体，称为L-适体，是一类有前途的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然很差，限制了这种方法在生物系统中应用的潜在效用。在本文中，我们使用新开发的“逆“体外选择方法对交叉手性L-适体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。 通过使用超过一百万个靶源序列的文库，我们确定了模型L-适体的RNA序列和结构偏好，并揭示了先前未鉴定的和潜在的广泛脱靶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，反向体外选择为推进L-适体技术提供了几个机会。
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 vitroselection 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, inversein vitroselection provides several opportunities to advance L‐aptamer technology.