Modular aptameric sensors.
We report the first examples of modular aptameric sensors, which transduce recognition events into fluorescence changes through allosteric regulation of noncovalent interactions with a fluorophore. These sensors consist of: (a) a reporting domain, which signals the binding event of an analyte through binding to a fluorophore; (b) a recognition domain, which binds the analyte; and (c) a communication module, which serves as a conduit between recognition and signaling domains. We tested recognition regions specific for ATP, FMN, and theophylline in combinations with malachite green binding aptamer as a signaling domain. In each case, we were able to obtain a functional sensor capable of responding to an increase in analyte concentration with an increase in fluorescence. Similar constructs that consist only of natural RNA could be expressed in cells and used as sensors for intracellular imaging.
Division of Experimental Therapeutics, Department of Medicine, Columbia University, New York, New York 10032, USA. mns18@columbia.edu
Milan N
MN
Stojanovic
Dmitry M
DM
Kolpashchikov
D000255:Adenosine Triphosphate / Q000032:analysis / Q000737:chemistry; D001483:Base Sequence; D015374:Biosensing Techniques / Q000379:methods*; D004396:Coloring Agents / Q000737:chemistry; D004247:DNA / Q000737:chemistry; D005486:Flavin Mononucleotide / Q000032:analysis; D005456:Fluorescent Dyes / Q000737:chemistry; D009690:Nucleic Acid Conformation; D009841:Oligonucleotides / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D012394:Rosaniline Dyes / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D013806:Theophylline / Q000032:analysis
D004396:Coloring Agents; D005456:Fluorescent Dyes; D009841:Oligonucleotides; D012394:Rosaniline Dyes; C005095:malachite green; D005486:Flavin Mononucleotide; D000255:Adenosine Triphosphate; D004247:DNA; D013806:Theophylline