title: In Vivo Production of Small Recombinant RNAs Embedded in a 5S rRNA-Derived Protective Scaffold.
abstract: Preparative synthesis of RNA is a challenging task that is usually accomplished using either chemical or enzymatic polymerization of ribonucleotides in vitro. Herein, we describe an alternative approach in which RNAs of interest are expressed as a fusion with a 5S rRNA-derived scaffold. The scaffold provides protection against cellular ribonucleases resulting in cellular accumulations comparable to those of regular ribosomal RNAs. After isolation of the chimeric RNA from the cells, the scaffold can be removed if necessary by deoxyribozyme-catalyzed cleavage followed by preparative electrophoretic separation of the cleavage reaction products. The protocol is designed for sustained production of high quality RNA on the milligram scale.
authors.affiliation: Department of Biology and Biochemistry, University of Houston, 3201 Cullen Blvd, Houston, TX, 77204-5001, USA, vstepanov@uh.edu.
authors.forename: Victor G
authors.initials: VG
authors.lastname: Stepanov
authors.forename: George E
authors.initials: GE
authors.lastname: Fox
mesh_terms: D021881:DNA, Catalytic / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D009690:Nucleic Acid Conformation; D010957:Plasmids / Q000235:genetics; D012341:RNA, Ribosomal, 5S / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification
chemical_list: D021881:DNA, Catalytic; D012341:RNA, Ribosomal, 5S