Detection of pre-mRNA splicing in vitro by an RNA-templated fluorogenic reaction.
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.
Nano Medical Engineering Laboratory, RIKEN Advanced Science Insutitute, 2-1 Hirosawa, Wako-Shi, Saitama 351-0198, Japan.
Yasutsugu
Y
Tamura
Kazuhiro
K
Furukawa
Rei
R
Yoshimoto
Yuto
Y
Kawai
Minoru
M
Yoshida
Satoshi
S
Tsuneda
Yoshihiro
Y
Ito
Hiroshi
H
Abe
D000818:Animals; D002645:Chickens; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes / Q000737:chemistry*; D012322:RNA Precursors / Q000032:analysis*; D012326:RNA Splicing; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D038207:delta-Crystallins / Q000235:genetics
D005456:Fluorescent Dyes; D012322:RNA Precursors; D012333:RNA, Messenger; D038207:delta-Crystallins