In vitro selection of signaling aptamers.
Reagentless biosensors that can directly transduce molecular recognition to optical signals should potentiate the development of sensor arrays for a wide variety of analytes. Nucleic acid aptamers that bind ligands tightly and specifically can be readily selected, but may prove difficult to adapt to biosensor applications. We have therefore attempted to develop selection methods that couple the broad molecular recognition properties of aptamers with signal transduction. Anti-adenosine aptamers were selected from a pool that was skewed to contain very few fluoresceinated uridines. The primary family of aptamers showed a doubling of relative fluorescence intensity at saturating concentrations of a cognate analyte, ATP, and could sense ATP concentrations as low as 25 microM. A single uridine was present in the best signaling aptamer. Surprisingly, other dyes could substitute for fluorescein and still specifically signal the presence of ATP, indicating that the single uridine functioned as a general "switch" for transducing molecular recognition to optical signals.
Jhaveri
S
S
Department of Chemistry and Biochemistry, University of Texas at Austin, Austin, TX 78712, USA.
Rajendran
M
M
Ellington
A D
AD
D000241:Adenosine / Q000737:chemistry; D000255:Adenosine Triphosphate / Q000378:metabolism; D001483:Base Sequence; D001665:Binding Sites; D015374:Biosensing Techniques; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D008024:Ligands; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D012313:RNA / Q000737:chemistry / Q000378:metabolism; D015398:Signal Transduction; D014529:Uridine / Q000737:chemistry
D005456:Fluorescent Dyes; D008024:Ligands; D012313:RNA; D000255:Adenosine Triphosphate; D000241:Adenosine; D014529:Uridine