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A viroid-derived system to produce large amounts of recombinant RNA in Escherichia coli.
Sci Rep. 2018 01 30; 8(1):1904.SR

Abstract

Viruses have been engineered into useful biotechnological tools for gene therapy or to induce the synthesis of products of interest, such as therapeutic proteins and vaccines, in animal and fungal cells, bacteria or plants. Viroids are a particular class of infectious agents of higher plants that exclusively consist of a small non-protein-coding circular RNA molecule. In the same way as viruses have been transformed into useful biotechnological devices, can viroids be converted into beneficial tools? We show herein that, by expressing Eggplant latent viroid (ELVd) derived RNAs in Escherichia coli together with the eggplant tRNA ligase, this being the enzyme involved in viroid circularization in the infected plant, RNAs of interest like aptamers, extended hairpins, or other structured RNAs are produced in amounts of tens of milligrams per liter of culture. Although ELVd fails to replicate in E. coli, ELVd precursors self-cleave through the embedded hammerhead ribozymes and the resulting monomers are, in part, circularized by the co-expressed enzyme. The mature viroid forms and the protein likely form a ribonucleoprotein complex that transitorily accumulates in E. coli cells at extraordinarily amounts.

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Authors+Show Affiliations

Daròs JA
Instituto de Biología Molecular y Celular de Plantas (Consejo Superior de Investigaciones Científicas-Universitat Politècnica de València), 46022, Valencia, Spain. jadaros@ibmcp.upv.es.
Aragonés V
Instituto de Biología Molecular y Celular de Plantas (Consejo Superior de Investigaciones Científicas-Universitat Politècnica de València), 46022, Valencia, Spain.
Cordero T
Instituto de Biología Molecular y Celular de Plantas (Consejo Superior de Investigaciones Científicas-Universitat Politècnica de València), 46022, Valencia, Spain.

MeSH

Escherichia coliPlant DiseasesRNARNA Ligase (ATP)RNA, CatalyticRNA, CircularRNA, ViralRibonucleoproteinsSolanum melongenaViroids

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

29382906

Citation

Daròs, José-Antonio, et al. "A Viroid-derived System to Produce Large Amounts of Recombinant RNA in Escherichia Coli." Scientific Reports, vol. 8, no. 1, 2018, p. 1904.
Daròs JA, Aragonés V, Cordero T. A viroid-derived system to produce large amounts of recombinant RNA in Escherichia coli. Sci Rep. 2018;8(1):1904.
Daròs, J. A., Aragonés, V., & Cordero, T. (2018). A viroid-derived system to produce large amounts of recombinant RNA in Escherichia coli. Scientific Reports, 8(1), 1904. https://doi.org/10.1038/s41598-018-20314-3
Daròs JA, Aragonés V, Cordero T. A Viroid-derived System to Produce Large Amounts of Recombinant RNA in Escherichia Coli. Sci Rep. 2018 01 30;8(1):1904. PubMed PMID: 29382906.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A viroid-derived system to produce large amounts of recombinant RNA in Escherichia coli. AU - Daròs,José-Antonio, AU - Aragonés,Verónica, AU - Cordero,Teresa, Y1 - 2018/01/30/ PY - 2017/08/09/received PY - 2018/01/17/accepted PY - 2018/2/1/entrez PY - 2018/2/1/pubmed PY - 2018/12/12/medline SP - 1904 EP - 1904 JF - Scientific reports JO - Sci Rep VL - 8 IS - 1 N2 - Viruses have been engineered into useful biotechnological tools for gene therapy or to induce the synthesis of products of interest, such as therapeutic proteins and vaccines, in animal and fungal cells, bacteria or plants. Viroids are a particular class of infectious agents of higher plants that exclusively consist of a small non-protein-coding circular RNA molecule. In the same way as viruses have been transformed into useful biotechnological devices, can viroids be converted into beneficial tools? We show herein that, by expressing Eggplant latent viroid (ELVd) derived RNAs in Escherichia coli together with the eggplant tRNA ligase, this being the enzyme involved in viroid circularization in the infected plant, RNAs of interest like aptamers, extended hairpins, or other structured RNAs are produced in amounts of tens of milligrams per liter of culture. Although ELVd fails to replicate in E. coli, ELVd precursors self-cleave through the embedded hammerhead ribozymes and the resulting monomers are, in part, circularized by the co-expressed enzyme. The mature viroid forms and the protein likely form a ribonucleoprotein complex that transitorily accumulates in E. coli cells at extraordinarily amounts. SN - 2045-2322 UR - https://www.unboundmedicine.com/medline/citation/29382906/A_viroid_derived_system_to_produce_large_amounts_of_recombinant_RNA_in_Escherichia_coli_ DB - PRIME DP - Unbound Medicine ER -