@article{APS6690,
author = {Carlos Fabián Flores-jasso and Catalina Arenas-huertero and Jose Luis Reyes and Cecilia Contreras-cubas and Alejandra Covarrubias and Luis Vaca},
title = {First step in pre-miRNAs processing by human Dicer},
journal = {Acta Pharmacologica Sinica},
volume = {30},
number = {8},
year = {2016},
keywords = {},
abstract = {Aim: To investigate the strand preference of the initial cleavage of human pre-miRNAs by human Dicer in vitro.
Methods: We used a series of in vitro transcribed pre-miRNAs that were radioactively labeled at their 5′ or 3′ ends in cleavage reactions with recombinant human Dicer or HeLa cytoplasmic S100 extracts. Pre-miRNAs samples were purified by denaturing and native PAGE and only the stem-loop structures were used in the experiments. Products of cleavage reactions were resolved by denaturing PAGE, and scanned by phosphor-imaging.
Results: Recombinant hDicer performs a biased first-cleavage in the pre-let-7b and hsa-pre-miR-17 3′ strand. This result is recapitulated in HeLa S100 cytoplasmic extracts.
Conclusion: The differential first-nick is observed in cleavage reactions only when stem-loops are substrates for hDicer.},
issn = {1745-7254}, url = {http://www.chinaphar.com/article/view/6690}
}