title: A novel strategy for selection of allosteric ribozymes yields RiboReporter sensors for caffeine and aspartame.
abstract: We have utilized in vitro selection technology to develop allosteric ribozyme sensors that are specific for the small molecule analytes caffeine or aspartame. Caffeine- or aspartame-responsive ribozymes were converted into fluorescence-based RiboReporter trade mark sensor systems that were able to detect caffeine or aspartame in solution over a concentration range from 0.5 to 5 mM. With read-times as short as 5 min, these caffeine- or aspartame-dependent ribozymes function as highly specific and facile molecular sensors. Interestingly, successful isolation of allosteric ribozymes for the analytes described here was enabled by a novel selection strategy that incorporated elements of both modular design and activity-based selection methods typically used for generation of catalytic nucleic acids.
authors.affiliation: Archemix Corporation, 1 Hampshire Street, Cambridge, MA 02139, USA.
authors.forename: Alicia
authors.initials: A
authors.lastname: Ferguson
authors.forename: Ryan M
authors.initials: RM
authors.lastname: Boomer
authors.forename: Markus
authors.initials: M
authors.lastname: Kurz
authors.forename: Sara C
authors.initials: SC
authors.lastname: Keene
authors.forename: John L
authors.initials: JL
authors.lastname: Diener
authors.forename: Anthony D
authors.initials: AD
authors.lastname: Keefe
authors.forename: Charles
authors.initials: C
authors.lastname: Wilson
authors.forename: Sharon T
authors.initials: ST
authors.lastname: Cload
mesh_terms: D000494:Allosteric Regulation; D001218:Aspartame / Q000032:analysis*; D001483:Base Sequence; D015374:Biosensing Techniques / Q000379:methods*; D002110:Caffeine / Q000032:analysis*; D019020:Directed Molecular Evolution; D008969:Molecular Sequence Data; D016337:RNA, Catalytic / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D013050:Spectrometry, Fluorescence
chemical_list: D016337:RNA, Catalytic; C427204:hammerhead ribozyme; D002110:Caffeine; D001218:Aspartame