A novel strategy for selection of allosteric ribozymes yields RiboReporter sensors for caffeine and aspartame.
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.
Ferguson
Alicia
A
Archemix Corporation, 1 Hampshire Street, Cambridge, MA 02139, USA.
Boomer
Ryan M
RM
Kurz
Markus
M
Keene
Sara C
SC
Diener
John L
JL
Keefe
Anthony D
AD
Wilson
Charles
C
Cload
Sharon T
ST
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
D016337:RNA, Catalytic; C427204:hammerhead ribozyme; D002110:Caffeine; D001218:Aspartame