siRNA design, production and delivery

描述（由申请人提供）：RNA干扰是一种新的强大的研究工具，可用于下调或敲除各种细胞类型和生物体中的特定基因。目前这项技术被用于研究基因功能，但有一天可能会用于人类治疗。RNAi现象最近才被证明存在于哺乳动物细胞中，并且在哺乳动物细胞中特异性地响应于称为小干扰RNA（siRNA）的短双链RNA分子而发生。在我们的研究中，我们发现靶向mRNA不同区域的siRNA分子在下调靶基因表达的能力方面存在显著差异。由于siRNA的作用机制是推测性的，我们不知道这种效力变化的原因。为了能够可靠地设计有效的siRNA，而不是使用昂贵的命中或失误的方法，必须确定现象背后的机制和siRNA设计的指导方针。我们建议进行详细和系统的分析，以阐明有效的siRNA设计的指导方针。作为这项研究的一部分，我们将通过对一组具有学术和临床重要性的基因产生siRNA分子来测试我们的指导方针，每一个基因都在一个明确的生物学途径中发挥作用。最后，我们将研究制造siRNA的方法以及使siRNA转染更有效的方法和试剂。根据本SBIR进行的研究将产生几种产品。首先，siRNA设计指南将用于设计高活性基因特异性siRNA，这些siRNA将作为对照、功能研究、治疗或诊断出售。我们还将出售用于siRNA酶促生产的试剂盒。这将显著降低siRNA合成的成本。最后，我们将销售允许siRNA在哺乳动物细胞中摄取和活性的转染剂。

DESCRIPTION (provided by applicant): RNA interference is a new and powerful research tool that can be used to down regulate or knock out a specific gene or genes in a variety of cell types and organisms. Currently this technology is being used to study gene function, but may one day be used for human therapeutics. The RNAi phenomenon was only recently demonstrated to be present in mammalian cells and in mammalian cells occur specifically in response to short double stranded RNA molecules termed small interfering RNAs (siRNA). In our research, we have discovered that siRNA molecules targeted to different regions of a mRNA vary strikingly in their ability to down regulate the expression of the target gene. Since the mechanism of action by which siRNA acts is speculative, we do not know the reason for this variability in potency. In order to be able to reliably design potent siRNA using something other than an expensive hit-or-miss approach, the mechanism behind the phenomenon and the guidelines of siRNA design must be determined. We propose to perform a detailed and systematic analysis to elucidate guidelines of effective siRNA design. As part of this research, we will test our guidelines by producing siRNA molecules to a panel of genes that have academic and clinical importance, each of which function in a well-defined biological pathway. Finally, we will investigate methods to make siRNAs and methods and reagents to make siRNA transfection more efficient. Several products will be generated from the research performed under this SBIR. First, the guidelines for siRNA design will be used to design highly active gene specific siRNAs that will be sold as controls, functional studies, therapeutics or diagnostics. We will also sell kits that will be used for the enzymatic production of siRNAs. This will reduce the cost of siRNA synthesis significantly. Lastly, we will sell transfection agents that allow for siRNA uptake and activity in mammalian cells.