Over the past five decades, DNA restriction enzymes have revolutionized biotechnology. While these enzymes are widely used in DNA research and DNA engineering, the emerging field of RNA and mRNA therapeutics requires sequence-specific RNA endoribonucleases. Here, we describe EcoToxN1, a member of the type III toxin-antitoxin family of sequence-specific RNA endoribonucleases, and its use in RNA and mRNA analysis. This enzyme recognizes a specific pentamer in a single-stranded RNA and cleaves the RNA within this sequence. The enzyme is neither dependent on annealing of guide RNA or DNA oligos to the template nor does it require magnesium. Furthermore, it performs over a wide range of temperatures. With its unique functions and characteristics, EcoToxN1 can be classified as an RNA restriction enzyme. EcoToxN1 enables new workflows in RNA analysis and biomanufacturing, meeting the demand for faster, cheaper, and more robust analysis methods.

中文翻译：

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使用溶核毒素作为限制性内切酶可实现新的 RNA 应用


在过去的五年里，DNA 限制性内切酶彻底改变了生物技术。虽然这些酶广泛用于 DNA 研究和 DNA 工程，但新兴的 RNA 和 mRNA 治疗领域需要序列特异性的 RNA 内切核糖核酸酶。在这里，我们描述了 EcoToxN1，它是序列特异性 RNA 核糖核酸内切酶的 III 型毒素-抗毒素家族的成员，及其在 RNA 和 mRNA 分析中的应用。该酶可识别单链 RNA 中的特定五聚体并切割该序列中的 RNA。该酶既不依赖于引导 RNA 或 DNA 寡核苷酸与模板的退火，也不需要镁。此外，它可以在很宽的温度范围内运行。凭借其独特的功能和特性，EcoToxN1 可归类为 RNA 限制性酶。 EcoToxN1 支持 RNA 分析和生物制造的新工作流程，满足对更快、更便宜和更强大的分析方法的需求。