Sequence-specific detection of MicroRNAs by signal-amplifying ribozymes.
The rational and straightforward design of hairpin ribozymes that can be sequence-specifically induced by external oligonucleotides is described. Due to intrinsic signal amplification, their sensitivity is at least an order of magnitude increased compared to standard molecular beacons. We applied this system to the detection of microRNAs, a recently discovered class of small endogenous RNA molecules that are involved in gene regulation. We show that the cognate microRNA can reliably and sensitively be detected at low concentrations in a mix of other microRNA sequences. These probes may be useful in applications that require direct detection of minute amounts of small DNAs or RNAs.
Hartig
Jörg S
JS
Kekulé Institut für Organische Chemie und Biochemie, Universität Bonn, Gerhard-Domagk-Strasse 1, 53121 Bonn, Germany.
Grüne
Imke
I
Najafi-Shoushtari
S Hani
SH
Famulok
Michael
M
D000818:Animals; D002384:Catalysis; D004331:Drosophila melanogaster / Q000235:genetics; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods; D035683:MicroRNAs / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D009690:Nucleic Acid Conformation; D016337:RNA, Catalytic / Q000737:chemistry* / Q000378:metabolism; D012680:Sensitivity and Specificity; D012815:Signal Processing, Computer-Assisted
D035683:MicroRNAs; D016337:RNA, Catalytic