Convenient rapid and portable tool for the detection of ribonucleases

总结/摘要 在过去的十年里，RNA的重要新商业应用不断增长。 RNA分子已成为基因表达的卓越调节因子。他们有一种独特的能力， 编辑基因组并破坏基因产物的稳定性。RNA的新的诊断应用正在不断地被开发出来。 发现了最后但并非最不重要的是，修饰的mRNA纳米颗粒制剂使得能够开发新的 疫苗和细胞疗法，其速度和功效是其他工艺无法比拟的。这个新发现的 成功已经在RNA研究的许多领域推动了增加的兴趣和努力。研究人员不断 发现RNA在调节细胞过程中的重要新作用。它也打开了大门， 为初创公司和大型制药和诊断公司提供了许多新的商业机会。 因此，现在大量的努力指向RNA的生产放大和大规模制造 分子。随着生产的增加，对制造业的关注和控制的需求大大增加 流程.对于RNA来说尤其如此，因为它很容易被污染的酶降解 在周围的环境中。 随着生产规模和成本的增加，必须更加注意严格的过程控制， 质量管理例如，用于受管制的治疗和诊断应用的RNA的制造 必须在经过认证的无RNA酶环境中进行。目前的商业方法，以验证不存在 需要昂贵的设备和数小时来进行。进行RNAse所需的成本和时间 测试阻碍了制造商和研究人员的生产力。一种新的快速工具来测试不需要的 RNA污染将极大地帮助研究人员和制造商，他们必须经常测试他们的产品。 环境和供应链材料。 在第一阶段，我们将创建一个新的原型侧向流测试的RNA酶的灵敏检测。我们 这项工作将产生一个测试包产品，与目前可用的工具相比，它具有几个重要的优点： 方便：我们的测试将是便携式的，不需要笨重，昂贵的实验室设备。灵敏度：我们的测试将 比现有方法灵敏至少5倍。时间：用户将在不到15分钟内看到结果 几分钟而不是几小时。我们提出的产品的这些综合优势将创造一个非常有效的 这是一个工具，以进一步支持RNA治疗和研究的持续创新。 具体目标#1：优化孵育缓冲液和RNA寡核苷酸序列，以改善RNA酶测定 灵敏度 具体目标#2：产生更灵敏的荧光版本的侧向流RNA酶检测测定。 具体目标#3：能够使用拭子擦拭物测试固体表面。

Summary/Abstract The past decade has brought an escalating growth of important new commercial applications for RNA. RNA molecules have emerged as the preeminent regulators of gene expression. They have a unique ability to edit genomes and destabilize gene products. New diagnostic applications for RNA are continually being discovered. Last but not least, modified mRNA nanoparticle formulations are enabling the development of new vaccines and cellular therapies with a speed and efficacy unrivaled by other processes. This newfound success has driven increased interest and efforts in many areas of RNA research. Researchers are constantly discovering important new roles for RNA in the regulation of cellular processes. It has also opened the door to many new commercial opportunities for startups and large pharmaceutical and diagnostic companies alike. Accordingly, massive effort is now directed to production scale-up and large-scale manufacturing of RNA molecules. As production increases, there is a far greater need for care and control of manufacturing processes. This is especially true for RNA since it is quite vulnerable to degradation by contaminating enzymes in the surrounding environment. As production scales and costs increase, more attention must be paid to strict process control and quality management. For example, manufacturing of RNA for regulated therapeutic and diagnostic applications must be performed in a certified RNAse-free environment. Current commercial methods to verify the absence of RNAse require expensive equipment and hours to perform. The cost and time required to perform RNAse testing hinders the productivity of manufacturers and researchers alike. A new rapid tool to test for unwanted RNA contamination would great assist researchers and manufacturers who must frequently test their environments and supply chain materials. In Phase I, we will create a novel prototype lateral flow test for the sensitive detection of RNAse. Our work will produce a test kit product with several important advantages over currently available tools: Convenience: Our test will be portable and not require bulky, expensive lab equipment. Sensitivity: Our test will be at least 5-times more sensitive than current methods. Time: Users will see their results in less than 15 minutes rather than hours. These combined advantages of our proposed product will create a highly effective tool to further support the continuing innovation in RNA therapeutics and research. Specific Aim #1: Optimize the incubation buffer and RNA oligonucleotide sequence to improve RNAse assay sensitivity. Specific Aim #2: Produce a more sensitive fluorescent version of the lateral flow RNAse detection assay. Specific Aim #3: Enable testing of solid surfaces using swab wipes.