Convenient rapid and portable tool for the detection of ribonucleases

用于检测核糖核酸酶的方便、快速、便携的工具

• 批准号: 10760552
• 负责人: LANCE P FORD
• 金额: $ 23万
• 依托单位: ATTOGENE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10760552
• 关键词: Accidents; Address; Area; Attention; Basic Science; Binding; Biological Assay; Biology; Buffers; COVID-19; Caring; Catalysis; Cell Therapy; Cell physiology; Certification; Communities; DNA; Data; Data Analyses; Detection; Detergents; Development; Devices; Diagnostic; Diagnostic Reagent Kits; Disease; Environment; Enzymes; Equipment; Fluorescence; Formulation; Freeze Drying; Gene Expression; Goals; Growth; High temperature of physical object; Hour; Incubated; Industry; Lateral; Liquid substance; Manufacturer; Messenger RNA; Methods; Modification; Monitor; Oligonucleotides; Pancreatic ribonuclease; Pharmacologic Substance; Phase; Procedures; Process; Production; Productivity; Quantum Dots; RNA; RNA Probes; RNA vaccine; Reagent; Regulator Genes; Research; Research Personnel; Resistance; Ribonucleases; Role; Sampling; Sodium Chloride; Solid; Speed; Streptavidin; Surface; Swab; System; Testing; Therapeutic; Time; Tube; Vaccine Production; Vaccine Therapy; Vaccines; Visualization; Work; cell growth regulation; commercial application; commercialization; cost; detection assay; experimental study; fighting; gene product; genome editing; improved; innovation; instrumentation; interest; laboratory equipment; laboratory facility; large scale production; lateral flow assay; manufacture; manufacturing facility; manufacturing process; manufacturing scale-up; nanoparticle; novel; novel diagnostics; novel vaccines; nuclease; particle; portability; preservation; prototype; rapid detection; real world application; success; supply chain; therapeutic RNA; tool; transcriptomics; vaccine immunotherapy

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.

摘要/摘要 过去的十年使RNA重要的新商业应用增长了增长。 RNA分子已成为基因表达的杰出调节剂。他们具有独特的能力 编辑基因组并破坏基因产品。 RNA的新诊断应用不断 发现。最后但并非最不重要的一点是，经过修改的mRNA纳米颗粒制剂使新的发展 疫苗和细胞疗法的速度和疗效与其他过程无与伦比。这个新发现 成功推动了RNA研究许多领域的兴趣和努力。研究人员一直在 发现RNA在细胞过程调节中的重要新作用。它也打开了 初创公司以及大型制药公司和诊断公司的许多新商业机会。 因此，现在大规模的努力用于生产规模和大规模制造RNA 分子。随着生产的增加，对制造业的护理和控制需要很大 过程。 RNA尤其如此，因为它很容易通过污染酶来降解 在周围的环境中。 随着生产规模和成本的增加，必须对严格的流程控制和 质量管理。例如，用于调节治疗和诊断应用的RNA制造 必须在经认证的无RNase环境中执行。当前验证缺席的商业方法 RNase需要昂贵的设备和工作时间才能执行。执行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率。一个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 环境和供应链材料。 在第一阶段，我们将为RNase敏感检测创建一个新型的原型侧流测试。我们的 与当前可用工具相比，工作将生产具有几个重要优势的测试套件产品： 便利：我们的测试将是可移植的，不需要笨重，昂贵的实验室设备。敏感性：我们的测试将 至少比当前方法更敏感。时间：用户将在不到15的时间内看到他们的结果 分钟而不是数小时。我们提出的产品的这些联合优势将创造出高效的 进一步支持RNA疗法和研究中持续创新的工具。 特定目标＃1：优化孵育缓冲液和RNA寡核苷酸序列以改善RNase分析 灵敏度。 特定的目标＃2：产生更灵敏的荧光版本的横向流动RNase检测测定法。 特定目标＃3：使用拭子擦除实体表面测试。