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

项目摘要/摘要 信使核糖核酸作为信使核糖核酸的载体，在细胞和生物体中发挥着重要作用。 蛋白质编码信息，并作为基因表达的调节。制药业已经 开始利用信使核糖核酸的许多天然功能来开发新的疗法和疫苗， 它们在预防和治疗疾病方面具有很高的有效性和极大的灵活性 从癌症和传染病(如肝炎和艾滋病毒)到遗传病 囊性纤维化和罕见疾病是由遗传基因缺陷引起的。 在信使核糖核酸疗法和信使核糖核酸疫苗方面的努力的扩大创造了对 大批量生产的信使核糖核酸分子的精确规格。尤其是，需要 产生长度为1kb的无不需要的副产物的信使核糖核酸分子 加入修饰的核苷酸，更有效的传递，更高的稳定性和更好的临床 药效，加剧了这个制造问题。尽管RNA可以被生产出来 在体外利用专门的RNA聚合酶进行酶促，这种酶被广泛用于 为研发目的生产RNA不适合适用于RNA的苛刻规格 用于基因治疗和疫苗的分子。一类新的酶，高度 为合成具有特定序列和结构的长RNA而优化，需要创建 满足这一新需求。 在第一阶段可行性项目中，Primorial Genetics发现并测试了53种新的RNA 其中13种聚合酶被发现优于目前用于RNA的酶 制造业。凭借我们第一阶段的业绩，该公司开始建立联系 对测试我们的新酶非常感兴趣的RNA治疗公司。 在拟议的第二阶段项目中，我们将改进其中四种酶，表征 6种改进的酶的活性的详细信息，并准备使用这些酶的方法和数据集 用于临床制造信使核糖核酸的酶。我们的目标是创造出能够满足不同需求的酶 需要制造不同的信使核糖核酸序列、大小和化学结构 在基因疫苗和正在开发中的基因治疗产品中具有代表性。这些酶 本工作中的发现和改进将直接用于mRNA的制造应用， 并将被许可或出售给开发mrna疫苗和疗法的公司以及 建设RNA制造能力的公司。

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.