Abstract

Genome-derived noncoding RNAs (ncRNAs), including microRNAs (miRNAs), small interfering RNAs (siRNAs), and long noncoding RNAs (lncRNAs), play an essential role in the control of target gene expression underlying various cellular processes, and dysregulation of ncRNAs is involved in the pathogenesis and progression of various diseases in virtually all species including humans. Understanding ncRNA biology has opened new avenues to develop novel RNA-based therapeutics. Presently, ncRNA research and drug development is dominated by the use of ncRNA mimics that are synthesized chemically in vitro and supplemented with extensive and various types of artificial modifications and thus may not necessarily recapitulate the properties of natural RNAs generated and folded in living cells in vivo. Therefore, there are growing interests in developing novel technologies for in vivo production of RNA molecules. The two most recent major breakthroughs in achieving an efficient, large-scale, and cost-effective fermentation production of recombinant or bioengineered RNAs (e.g., tens of milligrams from 1 L of bacterial culture) are (1) using stable RNA carriers and (2) direct overexpression in RNase III-deficient bacteria, while other approaches offer a low yield (e.g., nano- to microgram scales per liter). In this article, we highlight these novel microbial fermentation-based technologies that have shifted the paradigm to the production of true biological ncRNA molecules for research and development.

中文翻译：

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有效体内发酵生产非编码 RNA 的新方法。

摘要

基因组衍生的非编码 RNA (ncRNA)，包括 microRNA (miRNA)、小干扰 RNA (siRNA) 和长非编码 RNA (lncRNA)，在控制各种细胞过程中的靶基因表达和 ncRNA 失调中发挥着重要作用参与几乎所有物种（包括人类）的各种疾病的发病机制和进展。了解 ncRNA 生物学为开发新型 RNA 疗法开辟了新途径。目前，ncRNA 研究和药物开发主要以使用 ncRNA 模拟物为主，这些模拟物在体外化学合成，并辅以广泛和各种类型的人工修饰，因此不一定能重现活细胞体内生成和折叠的天然 RNA 的特性。因此，人们对开发用于体内生产RNA分子的新技术越来越感兴趣。在实现重组或生物工程 RNA（例如，从 1 L 细菌培养物中生产数十毫克）的高效、大规模和经济有效的发酵生产方面，最近取得的两项重大突破是 (1) 使用稳定的 RNA 载体和 (2 ）在缺乏 RNase III 的细菌中直接过度表达，而其他方法产量较低（例如，每升纳微克级）。在本文中，我们重点介绍这些基于微生物发酵的新型技术，这些技术已将范式转变为生产真正的生物 ncRNA 分子以供研究和开发。