title: FRET-based optical assay for selection of artificial riboswitches.
abstract: Artificial riboswitches are engineered to regulate gene expression in response to a variety of non-endogenous small molecules and, therefore, can be useful tools to reprogram cellular behavior for different applications. A new synthetic riboswitch can be created by linking an in vitro-selected aptamer with a randomized expression platform followed by in vivo selection and screening. Here, we describe an in vivo selection and screening technique to discover artificial riboswitches in E. coli cells that is based on TEV protease-FRET substrate reporter system.
authors.affiliation: The Henry M. Jackson Foundation, Bethesda, MD, USA.
authors.forename: Svetlana V
authors.initials: SV
authors.lastname: Harbaugh
authors.forename: Molly E
authors.initials: ME
authors.lastname: Chapleau
authors.forename: Yaroslav G
authors.initials: YG
authors.lastname: Chushak
authors.forename: Morley O
authors.initials: MO
authors.lastname: Stone
authors.forename: Nancy
authors.initials: N
authors.lastname: Kelley-Loughnane
mesh_terms: D052157:Aptamers, Nucleotide / Q000235:genetics; D001426:Bacterial Proteins / Q000032:analysis / Q000235:genetics; D001483:Base Sequence; D010450:Endopeptidases / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D015870:Gene Expression; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D049408:Luminescent Agents / Q000032:analysis / Q000378:metabolism; D008164:Luminescent Proteins / Q000032:analysis / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D058928:Riboswitch
chemical_list: D052157:Aptamers, Nucleotide; D001426:Bacterial Proteins; D049408:Luminescent Agents; D008164:Luminescent Proteins; D058928:Riboswitch; C054562:yellow fluorescent protein, Bacteria; D049452:Green Fluorescent Proteins; D010450:Endopeptidases; C099488:TEV protease