title: New insights for therapeutic recombinant human miRNAs heterologous production: Rhodovolum sulfidophilum vs Escherichia coli.
abstract: 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.
authors.affiliation: a CICS-UBI - Health Sciences Research Centre, Universidade da Beira Interior, Avenida Infante D. Henrique , Covilhã , Portugal.
authors.forename: Patrícia
authors.initials: P
authors.lastname: Pereira
authors.affiliation: a CICS-UBI - Health Sciences Research Centre, Universidade da Beira Interior, Avenida Infante D. Henrique , Covilhã , Portugal.
authors.forename: Augusto Q
authors.initials: AQ
authors.lastname: Pedro
authors.affiliation: a CICS-UBI - Health Sciences Research Centre, Universidade da Beira Interior, Avenida Infante D. Henrique , Covilhã , Portugal.
authors.forename: João A
authors.initials: JA
authors.lastname: Queiroz
authors.affiliation: b Faculty of Pharmacy, University of Coimbra, Azinhaga Sta. Comba , Coimbra , Portugal.|c REQUIMTE/LAQV, Group of Pharmaceutical Technology, Faculty of Pharmacy, University of Coimbra , Coimbra , Portugal.
authors.forename: Ana R
authors.initials: AR
authors.lastname: Figueiras
authors.affiliation: a CICS-UBI - Health Sciences Research Centre, Universidade da Beira Interior, Avenida Infante D. Henrique , Covilhã , Portugal.
authors.forename: Fani
authors.initials: F
authors.lastname: Sousa
mesh_terms: D019149:Bioreactors / Q000382:microbiology*; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics; D006801:Humans; D035683:MicroRNAs / Q000096:biosynthesis* / Q000235:genetics*; D011995:Recombination, Genetic / Q000235:genetics*; D041903:Rhodovulum / Q000235:genetics* / Q000378:metabolism
chemical_list: D035683:MicroRNAs
keywords: Escherichia coli; RNAi technology; Rhodovolum sulfidophilum; fermentation; miRNA; non-coding RNA; recombinant RNA