Development of superior polymerases for next-generation mRNA therapeutic & vaccine manufacturing

项目总结/摘要 信使核糖核酸（mRNA）在细胞和生物体中起着关键作用，作为细胞和生物体的载体。 蛋白质编码信息和作为基因表达的调节剂。医药行业 开始利用mRNA的许多天然功能来开发新的疗法和疫苗， 其在预防和治疗疾病方面具有高功效和极大的灵活性 从癌症和传染病，如肝炎和艾滋病毒，到遗传疾病， 囊性纤维化和遗传缺陷引起的罕见疾病。 mRNA治疗和mRNA疫苗的扩大努力已经创造了新的需求， mRNA分子被大量生产以达到精确的规格。特别是需要 产生长度> 1 kb的mRNA分子，其不含不需要的副产物，以及 掺入修饰核苷酸以更有效地递送、更高的稳定性和更好的临床应用 效率，使该制造问题复杂化。虽然RNA可以被生产出来， 使用专门的RNA聚合酶在体外进行酶促反应，所述酶广泛用于 为研发目的生产RNA不适合适用于RNA的苛刻规格 用于mRNA治疗剂和疫苗的分子。一种新型的酶， 优化合成具有特定序列和结构的长RNA，需要创建， 满足这一新需求。 在第一阶段的可行性项目中，原始遗传学公司发现并测试了53种新的RNA， 其中13种聚合酶被发现上级目前用于RNA聚合的酶。 制造业凭借我们第一阶段的成果，公司开始建立联系， 他们对测试我们的新酶非常感兴趣。 在拟议的第二阶段项目中，我们将改进其中的四种酶， 详细描述六种改进的酶的活性，并准备使用这些酶的方法和数据集。 用于临床mRNA制造的酶。我们的目标是创造酶，可以满足各种 需要制造多种mRNA序列、大小和化学结构 在mRNA疫苗和mRNA治疗产品的开发中。的酶 在这项工作中发现和改进的将直接用于mRNA制造应用， 并将被授权或出售给开发mRNA疫苗和治疗药物的公司， 公司建立RNA制造能力。

Project Summary/Abstract Messenger ribonucleic acid (mRNA) plays key roles in cells and organisms as a carrier of protein-coding information and as a regulator of gene expression. The pharmaceutical industry has begun to exploit the many natural functions of mRNA to develop novel therapeutics and vaccines, which promise high efficacy and great flexibility in the prevention and treatment of diseases ranging from cancers and infectious diseases such as hepatitis and HIV to genetic diseases such as cystic fibrosis and rare diseases caused by heritable genetic defects. The expanded effort in mRNA therapeutics and mRNA vaccines has created a new demand for mRNA molecules manufactured in large quantities to precise specifications. In particular, the need to create mRNA molecules >1kb in length that are free of unwanted side products, and to incorporate modified nucleotides for more efficient delivery, higher stability and better clinical efficacy, has compounded this manufacturing problem. Although RNAs can be produced enzymatically in vitro with the use of specialized RNA polymerases, the enzymes widely used to produce RNA for R&D purposes are not suited for the demanding specifications that apply to RNA molecules intended for mRNA therapeutics and vaccines. A new class of enzymes, highly optimized for synthesis of long RNAs with specific sequences and structures, need to be created to meet this new demand. In a Phase I feasibility project, Primordial Genetics discovered and tested 53 novel RNA polymerases of which 13 were found to be superior to the current enzymes used for RNA manufacturing. On the strength of our Phase I results, the company began to forge connections to RNA therapeutics companies who are very interested in testing our new enzymes. In the proposed Phase II project, we will improve four of these enzymes, characterize the activity of six improved enzymes in detail, and prepare methods and datasets for using these enzymes for clinical mRNA manufacturing. Our goal is to create enzymes that can meet the varied needs for manufacturing a diversity of mRNA sequences, sizes and chemical structures represented in mRNA vaccines and mRNA therapeutic products under development. The enzymes discovered and improved in this work will be directly useful for mRNA manufacturing applications, and will be licensed or sold to companies developing mRNA vaccines and therapeutics as well as companies building RNA manufacturing capabilities.