Fluorescence-based high-throughput screening of dicer cleavage activity

0434676 - ÚMG 2015 RIV US eng J - Článek v odborném periodiku
Podolská, Kateřina - Sedlák, David - Bartůněk, Petr - Svoboda, Petr
Fluorescence-based high-throughput screening of dicer cleavage activity.
Journal of Biomolecular Screening. Roč. 19, č. 3 (2014), s. 417-426. ISSN 1087-0571
Grant CEP: GA ČR GA13-29531S; GA MŠMT(CZ) LC06077; GA MŠMT LM2011022
Grant ostatní: EMBO(DE) 1483
Institucionální podpora: RVO:68378050
Klíčová slova: Dicer * siRNA * high-throughput screening
Kód oboru RIV: EB - Genetika a molekulární biologie
Impakt faktor: 2.423, rok: 2014

Production of small RNAs by ribonuclease III Dicer is a key step in microRNA and RNA interference pathways, which employ Dicer-produced small RNAs as sequence-specific silencing guides. Further studies and manipulations of microRNA and RNA interference pathways would benefit from identification of small-molecule modulators. Here, we report a study of a fluorescence-based in vitro Dicer cleavage assay, which was adapted for high-throughput screening. The kinetic assay can be performed under single-turnover conditions (35 nM substrate and 70 nM Dicer) in a small volume (5 µL), which makes it suitable for high-throughput screening in a 1536-well format. As a proof of principle, a small library of bioactive compounds was analyzed, demonstrating potential of the assay.
Trvalý link: http://hdl.handle.net/11104/0241985