Detection of pre-mRNA splicing in vitro by an RNA-templated fluorogenic reaction

• Published in: Bioorganic & medicinal chemistry letters Vol. 22; no. 23; pp. 7248 - 7251
• Main Authors: Tamura, Yasutsugu, Furukawa, Kazuhiro, Yoshimoto, Rei, Kawai, Yuto, Yoshida, Minoru, Tsuneda, Satoshi, Ito, Yoshihiro, Abe, Hiroshi
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier Ltd 01-12-2012; Elsevier
• Subjects: Animals; Biological and medical sciences; CDC gene; chemistry; Chickens; delta-Crystallins - genetics; DNA probes; Drug discovery; drugs; Fluorescence; Fluorescence Resonance Energy Transfer; Fluorescent Dyes - chemistry; genes; Medical sciences; messenger RNA; methodology; mRNA; Oligonucleotides; Pharmacology. Drug treatments; RNA; RNA Precursors - analysis; RNA Splicing; RNA, Messenger - genetics; RNA, Messenger - metabolism; RNA-templated reaction; Splicing; Fluorescence; RNA-templated reaction; RNA; Splicing; Messenger RNA; Detection; In vitro
• ISSN: 0960-894X; 1464-3405
• DOI: 10.1016/j.bmcl.2012.09.033

RNA splicing is an important target for basic research of disease mechanisms and for drug discovery. Here, we report a new method for analysis of the in vitro RNA splicing process that produces fluorescence using a reduction-triggered fluorescence (RETF) probe. The fluorescence signal is produced only when the two probes bind side-by-side with a specific RNA target. Precursor messenger RNA and mature messenger RNA originating from the chicken δ-crystallin (CDC) gene were successfully discriminated in solution using an RETF probe with the assistance of helper oligonucleotide strands. Also, we successfully applied RETF probes to the detection of emerging mature mRNA in an in vitro splicing process.