New insights for therapeutic recombinant human miRNAs heterologous production: Rhodovolum sulfidophilum vs Escherichia coli

Bioengineered

João A. Queiroz¹  Fani Sousa¹  Augusto Q. Pedro¹  Patrícia Pereira¹  Ana R. Figueiras² 

[1] CICS-UBI – Health Sciences Research Centre, Universidade da Beira Interior, Avenida Infante D. Henrique, Covilhã, Portuga;Faculty of Pharmacy, University of Coimbra, Azinhaga Sta. Comba, Coimbra, Portuga;REQUIMTE/LAQV, Group of Pharmaceutical Technology, Faculty of Pharmacy, University of Coimbra, Coimbra, Portuga;

关键词: Escherichia coli;    fermentation;    miRNA;    non-coding RNA;    recombinant RNA;    RNAi technology;    Rhodovolum sulfidophilum;   

DOI  :  10.1080/21655979.2017.1284710

来源: Taylor & Francis

【 摘 要 】

RNA interference-based technologies have emerged as an attractive and effective therapeutic option with potential application in diverse human diseases. These tools rely on the development of efficient strategies to obtain homogeneous non-coding RNA samples with adequate integrity and purity, thus avoiding non-targeted gene-silencing and related side-effects that impair their application onto pre-clinical practice. These RNAs have been preferentially obtained by in vitro transcription using DNA templates or via chemical synthesis. As an alternative to overcome the limitations presented by these methods, in vivo recombinant production of RNA biomolecules has become the focus in RNA synthesis research. Therefore, using pre-miR-29b as a model, here it is evaluated the time-course profile of Escherichia coli and Rhodovolum sulfidophilum microfactories to produce this microRNA. As the presence of major host contaminants arising from the biosynthesis process may have important implications in the subsequent downstream processing, it is also evaluated the production of genomic DNA and host total proteins. Considering the rapidly growing interest on these innovative biopharmaceuticals, novel, more cost-effective, simple and easily scaled-up technologies are highly desirable. As microRNA recombinant expression fulfills those requirements, it may take the leading edge in the methodologies currently available to obtain microRNAs for clinical or structural studies.

【 授权许可】

Unknown