Immobilized RNA switches for the analysis of complex chemical and biological mixtures.
A prototype biosensor array has been assembled from engineered RNA molecular switches that undergo ribozyme-mediated self-cleavage when triggered by specific effectors. Each type of switch is prepared with a 5'-thiotriphosphate moiety that permits immobilization on gold to form individually addressable pixels. The ribozymes comprising each pixel become active only when presented with their corresponding effector, such that each type of switch serves as a specific analyte sensor. An addressed array created with seven different RNA switches was used to report the status of targets in complex mixtures containing metal ion, enzyme cofactor, metabolite, and drug analytes. The RNA switch array also was used to determine the phenotypes of Escherichia coli strains for adenylate cyclase function by detecting naturally produced 3',5'- cyclic adenosine monophosphate (cAMP) in bacterial culture media.
Department of Molecular, Cellular and Developmental Biology, Yale University, New Haven, CT 06520-8103, USA.
S
S
Seetharaman
M
M
Zivarts
N
N
Sudarsan
R R
RR
Breaker
D000262:Adenylyl Cyclases / Q000378:metabolism; D000495:Allosteric Site; D015374:Biosensing Techniques / Q000379:methods*; D000242:Cyclic AMP / Q000378:metabolism; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004926:Escherichia coli / Q000378:metabolism; D005821:Genetic Techniques; D006046:Gold / Q000737:chemistry; D007700:Kinetics; D009690:Nucleic Acid Conformation; D009696:Nucleic Acids / Q000737:chemistry*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D010641:Phenotype; D012313:RNA / Q000737:chemistry* / Q000378:metabolism*; D016337:RNA, Catalytic / Q000378:metabolism; D012825:Silicon / Q000737:chemistry; D013997:Time Factors
D009696:Nucleic Acids; D016337:RNA, Catalytic; D012313:RNA; D006046:Gold; D004247:DNA; D000242:Cyclic AMP; D000262:Adenylyl Cyclases; D012825:Silicon