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Production of Recombinant RNA in Escherichia coli Using Eggplant Latent Viroid as a Scaffold.
Methods Mol Biol. 2022; 2316:315-327.MM

Abstract

Similar to viruses, viroids can also be engineered and transformed into useful biotechnological tools. We describe here a viroid-based system to produce large amounts of recombinant RNA in Escherichia coli. A precursor of eggplant latent viroid (ELVd), with the RNA of interest inserted between positions U245 and U246, is co-expressed in E. coli along the chloroplastic isoform of the eggplant tRNA ligase, the enzyme that mediates the circularization of this viroid in the infected plants. In the bacterial cells, the chimeric ELVd-RNA-of-interest precursor self-cleaves through the embedded hammerhead ribozymes, and the monomer is recognized and circularized by the co-expressed tRNA ligase. The resulting circular RNA, likely bound to the tRNA ligase, accumulates to a high concentration in the bacterial cells.

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Publisher Full Text (DOI)

Authors+Show Affiliations

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

MeSH

Escherichia coliNucleic Acid ConformationRNARNA Ligase (ATP)RNA, CatalyticRNA, ViralSolanum melongenaViroids

Pub Type(s)

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

Language

eng

PubMed ID

34845704

Citation

Ortolá, Beltrán, and José-Antonio Daròs. "Production of Recombinant RNA in Escherichia Coli Using Eggplant Latent Viroid as a Scaffold." Methods in Molecular Biology (Clifton, N.J.), vol. 2316, 2022, pp. 315-327.
Ortolá B, Daròs JA. Production of Recombinant RNA in Escherichia coli Using Eggplant Latent Viroid as a Scaffold. Methods Mol Biol. 2022;2316:315-327.
Ortolá, B., & Daròs, J. A. (2022). Production of Recombinant RNA in Escherichia coli Using Eggplant Latent Viroid as a Scaffold. Methods in Molecular Biology (Clifton, N.J.), 2316, 315-327. https://doi.org/10.1007/978-1-0716-1464-8_25
Ortolá B, Daròs JA. Production of Recombinant RNA in Escherichia Coli Using Eggplant Latent Viroid as a Scaffold. Methods Mol Biol. 2022;2316:315-327. PubMed PMID: 34845704.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Production of Recombinant RNA in Escherichia coli Using Eggplant Latent Viroid as a Scaffold. AU - Ortolá,Beltrán, AU - Daròs,José-Antonio, PY - 2021/11/30/entrez PY - 2021/12/1/pubmed PY - 2022/1/19/medline KW - Circular RNA KW - Escherichia coli KW - Hammerhead ribozyme KW - RNA ligation KW - Recombinant RNA KW - Ribonucleoprotein complex KW - tRNA ligase SP - 315 EP - 327 JF - Methods in molecular biology (Clifton, N.J.) JO - Methods Mol Biol VL - 2316 N2 - Similar to viruses, viroids can also be engineered and transformed into useful biotechnological tools. We describe here a viroid-based system to produce large amounts of recombinant RNA in Escherichia coli. A precursor of eggplant latent viroid (ELVd), with the RNA of interest inserted between positions U245 and U246, is co-expressed in E. coli along the chloroplastic isoform of the eggplant tRNA ligase, the enzyme that mediates the circularization of this viroid in the infected plants. In the bacterial cells, the chimeric ELVd-RNA-of-interest precursor self-cleaves through the embedded hammerhead ribozymes, and the monomer is recognized and circularized by the co-expressed tRNA ligase. The resulting circular RNA, likely bound to the tRNA ligase, accumulates to a high concentration in the bacterial cells. SN - 1940-6029 UR - https://www.unboundmedicine.com/medline/citation/34845704/Production_of_Recombinant_RNA_in_Escherichia_coli_Using_Eggplant_Latent_Viroid_as_a_Scaffold_ DB - PRIME DP - Unbound Medicine ER -