da Silva Nonato Nyelson, Nunes Leandro Silva, da Silveira Martins Amanda Weege, Pinhal Danillo, Domingues William Borges, Bellido-Quispe Dionet Keny, Remião Mariana Härter, Campos Vinicius Farias
Laboratório de Genômica Estrutural, Programa de Pós-Graduação em Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
Laboratório Genômica e Evolução Molecular, Instituto de Biociências de Botucatu, Departamento de Genética, Universidade Estadual Paulista, Botucatu, SP, Brazil.
Plasmid. 2024 Sep-Nov;131-132:102731. doi: 10.1016/j.plasmid.2024.102731. Epub 2024 Sep 28.
Bacteria, the primary microorganisms used for industrial molecule production, do not naturally generate miRNAs. This study aims to systematically review current literature on miRNA expression systems in bacteria and address three key questions: (1) Which microorganism is most efficient for heterologous miRNA production? (2) What essential elements should be included in a plasmid construction to optimize miRNA expression? (3) Which commercial plasmid is most used for miRNA expression? Initially, 832 studies were identified across three databases, with fifteen included in this review. Three species-Escherichia coli, Salmonella typhimurium, and Rhodovulum sulfidophilum-were found as host organisms for recombinant miRNA expression. A total of 78 miRNAs were identified, out of which 75 were produced in E. coli, one in R. sulfidophilum (miR-29b), and two in S. typhimurium (mi-INHA and miRNA CCL22). Among gram-negative bacteria, R. sulfidophilum emerged as an efficient platform for heterologous production, thanks to features like nucleic acid secretion, RNase non-secretion, and seawater cultivation capability. However, E. coli remains the widely recognized model for large-scale miRNA production in biotechnology market. Regarding plasmids for miRNA expression in bacteria, those with an lpp promoter and multiple cloning sites appear to be the most suitable due to their robust expression cassette. The reengineering of recombinant constructs holds potential, as improvements in construct characteristics maximize the expression of desired molecules. The utilization of recombinant bacteria as platforms for producing new molecules is a widely used approach, with a focus on miRNAs expression for therapeutic contexts.
细菌是用于工业分子生产的主要微生物，但它们不会自然产生微小RNA（miRNA）。本研究旨在系统回顾当前关于细菌中miRNA表达系统的文献，并回答三个关键问题：（1）哪种微生物最适合用于异源miRNA生产？（2）质粒构建中应包含哪些基本元件以优化miRNA表达？（3）哪种商业质粒最常用于miRNA表达？最初，在三个数据库中鉴定出832项研究，本综述纳入了其中15项。发现三种细菌——大肠杆菌、鼠伤寒沙门氏菌和嗜硫小红卵菌——作为重组miRNA表达的宿主生物体。总共鉴定出78种miRNA，其中75种在大肠杆菌中产生，1种在嗜硫小红卵菌中产生（miR-29b），2种在鼠伤寒沙门氏菌中产生（mi-INHA和miRNA CCL22）。在革兰氏阴性菌中，嗜硫小红卵菌由于具有核酸分泌、不分泌核糖核酸酶和海水培养能力等特性，成为异源生产的有效平台。然而，大肠杆菌仍然是生物技术市场上大规模生产miRNA的广泛认可的模型。关于细菌中miRNA表达的质粒，具有脂蛋白启动子和多克隆位点的质粒似乎是最合适的，因为它们具有强大的表达盒。重组构建体的改造具有潜力，因为构建体特性的改善可使所需分子的表达最大化。利用重组细菌作为生产新分子的平台是一种广泛使用的方法，重点是在治疗背景下表达miRNA。
