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文库自我选择优化基因特异性抑制剂的新方法。Lasso是与靶RNA分子杂交并拓扑连接的RNA分子，与目前可用的反义化合物相比，产生具有上级稳定性和特异性的复合物。RNA Lassos可用于抑制基因表达或作为探针检测与疾病相关的特定多核苷酸序列的存在。所有反义方法的主要挑战是在靶mRNA上找到“抗敏感”序列。我们的方法允许自动选择可访问的靶位点和最佳Lasso序列。我们描述了RNA Lasso文库的设计，该文库将基因特异性的、定向的反义文库沿着部分随机化的Lasso序列。将选择在不存在靶时为线性的Lasso，但在与靶序列结合时被激活以进行自连接，产生强的拓扑连接的复合物。然后测试这些选择的Lasso抑制靶mRNA翻译的能力。通过这种方式，优化的Lassos将以自动和直接的方式导出，用于作为靶标验证、基因功能分析和治疗剂的应用。

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.