Aptazyme-based riboswitches as label-free and detector-free sensors for cofactors.
We constructed a label-free and detector-free aptazyme-based riboswitch sensor for detecting the cofactor of the aptazyme. This riboswitch, which usually suppresses the gene expression with its anti-RBS sequence bound to the RBS of its own mRNA (OFF), activates the translation only when a cofactor is added to release the anti-RBS sequence from itself as a result of cofactor-induced self-cleavage by the aptazyme (ON). The rationally optimized one with beta-galactosidase as a reporter gene enabled us to detect the cofactor of the aptazyme visibly with high ON/OFF efficiency.
Ogawa
Atsushi
A
Bioengineering Laboratory, RIKEN (The Institute of Physical and Chemical Research), 2-1 Hirosawa, Wako, Saitama 351-0198, Japan. a-ogawa@riken.jp
Maeda
Mizuo
M
D052157:Aptamers, Nucleotide; D001483:Base Sequence; D001665:Binding Sites; D015374:Biosensing Techniques; D003067:Coenzymes / Q000032:analysis*; D004247:DNA / Q000737:chemistry; D015870:Gene Expression / Q000187:drug effects; D017930:Genes, Reporter; D008156:Luciferases / Q000235:genetics; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D014176:Protein Biosynthesis / Q000187:drug effects* / Q000235:genetics; D016337:RNA, Catalytic / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology*; D012333:RNA, Messenger / Q000737:chemistry; D012270:Ribosomes / Q000737:chemistry
D052157:Aptamers, Nucleotide; D003067:Coenzymes; D016337:RNA, Catalytic; D012333:RNA, Messenger; D004247:DNA; D008156:Luciferases