METHOD FOR DEVELOPING IMPROVED GENE INHIBITORS

本提案的目标是开发一种通用的方法来快速获得任何感兴趣的基因的有效抑制剂。实现这一目标需要两个步骤：1)在mRNA上选择一个可访问的位点，2)设计一个有效的分子来识别该位点并与之结合，或者通过其他方式使其失活。目前可用的反义技术和可访问的靶点选择还处于起步阶段，不可靠。因此，我们提出了一种新的通用方法，用于快速开发一种有效的技术，用于绘制任何RNA中的可达靶点，以及对这些可达靶点进行有效的工程抑制剂。我们通过使用反义序列的随机文库来实现这一目标，这些序列在靶RNA处于自然构象的细胞内起作用，自动识别任何所需靶RNA中最易接近的位点。通过使用这些已确定的可访问位点，我们将设计我们专有的，基于拓扑的反义设计，RNA挂锁，用于靶向抑制任何所需基因的mRNA。建议的商业应用：反义技术目前受到寡核苷酸与目标mrna上大多数位点结合能力差以及识别它们可以结合的位点的困难的阻碍。通过克服这一障碍，我们将有助于实现反义技术最初承诺的合理药物设计和药物靶点验证的巨大可能性。

The goal of this proposal is to develop a universal method for rapidly deriving effective inhibitors for any gene of interest. Accomplishing this goal requires two steps: 1) selecting an accessible site on the mRNA, and 2) designing an effective molecule that will recognize that site and bind to it, or inactivate it by some other means. Currently available antisense technology and accessible target site selection is still in its infancy and unreliable. Therefore we propose a novel and general approach for rapidly developing an effective technology for mapping accessible target sites in any RNA as well as engineering effective inhibitors to those mapped accessible sites. We accomplish this by using random libraries of antisense sequences that, functioning inside the cell where the target is in its natural conformation, automatically identify the most accessible sites in any desired target RNA. By employing those identified accessible sites, we will engineer our proprietary, topology-based antisense design, RNA Padlocks, for targeted inhibition of mRNA of any desired gene. PROPOSED COMMERCIAL APPLICATIONS: Antisense technologies are currently hindered by the poor ability of oligonucleotides to bind to most sites on target mRNAs, and the difficulty of identifying those sites to which they can bind. By overcoming this hurdle, we will help realize the great possibilities of rational drug design and drug target validation that antisense technology originally promised.