Library-based selection of optimal gene inhibitors

描述(由申请人提供)：我们提出了一种基于RNA Lasso文库的优化基因特异性抑制剂自我选择的新方法。Lassos是一种与靶RNA分子杂交并与其形成拓扑连接的RNA分子，与目前可用的反义化合物相比，它产生了具有更高稳定性和特异性的复合体。RNA Lassos可以用来抑制基因的表达，也可以用来作为探针来检测与疾病相关的特定多核苷酸序列的存在。在所有反义方法中，一个主要的挑战是在目标mRNAs上寻找“反敏感”序列。我们的方法允许自动选择可访问的目标位置和最佳套索序列。我们描述了RNA Lasso文库的设计，该文库结合了基因特异性的、定向的反义文库以及部分随机的Lasso序列。将选择在没有靶的情况下呈线性的套索，但在与靶序列结合时被激活以进行自我连接，从而产生强的、拓扑连接的复合体。然后将测试这些选定的Lassos是否具有抑制目标mRNA翻译的能力。这样，优化的Lassos将以自动和直接的方式获得，用于靶标验证、基因功能分析和治疗学的试剂。

DESCRIPTION (provided by applicant): We propose a new approach for the self-selection of optimized gene-specific inhibitors based on RNA Lasso libraries. Lassos are RNA molecules that hybridize to and become topologically linked with target RNA molecules, creating complexes with superior stability and specificity as compared to currently available antisense compounds. RNA Lassos can be employed either to inhibit the expression of genes or as probes to detect the presence of particular polynucleotide sequences associated with a disease. A major challenge in all antisense approaches is finding "antisensitive" sequences on target mRNAs. Our approach allows automatic selection of both accessible target sites and optimal Lasso sequences. We describe the design of RNA Lasso libraries that incorporate gene-specific, directed antisense libraries along with a partially randomized Lasso sequence. Lassos will be selected that are linear in the absence of the target, but are activated to undergo self-ligation upon binding to a target sequence, creating a strong, topologically linked complex. These selected Lassos will be then tested for their ability to inhibit translation of the target mRNA. In this way, optimized Lassos will be derived in an automatic and straightforward manner for application as agents for target validation, gene function analysis, and therapeutics.