title: Rapid Selection of RNA Aptamers that Activate Fluorescence of Small Molecules.
abstract: RNA aptamers can serve as valuable tools for studying and manipulating live cells. Fluorescent aptamers are the ones that bind to and turn on fluorescence of small-molecule dyes (fluorogens). Similarly to fluorescent proteins, fluorescent RNA aptamers can be used to image spatial and temporal RNA dynamics in live cells. Additionally, these aptamers can serve as a basis for engineering genetically encoded fluorescent biosensors. This chapter presents a protocol for rapid and efficient screening of RNA aptamer libraries to isolate fluorescent aptamers. The protocol describes how to design, clone, and express RNA aptamer library in bacterial cells and how to screen the bacteria to find aptamers with the desired fluorescent properties.
authors.affiliation: Essen Bioscience, Ann Arbor, MI, USA. grigory.filonov@essenbio.com.|Department of Pharmacology, Weill Medical College, Cornell University, New York, NY, USA. grigory.filonov@essenbio.com.
authors.forename: Grigory S
authors.initials: GS
authors.lastname: Filonov
mesh_terms: D052157:Aptamers, Nucleotide / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D005456:Fluorescent Dyes / Q000737:chemistry; D015723:Gene Library; D006801:Humans; D052156:SELEX Aptamer Technique; D054852:Small Molecule Libraries / Q000737:chemistry* / Q000378:metabolism
chemical_list: D052157:Aptamers, Nucleotide; D005456:Fluorescent Dyes; D054852:Small Molecule Libraries
keywords: Aptamer; Directed evolution; FACS; Fluorescence; High-throughput screening; RNA