In-vivo quantification of primary microRNA processing by Drosha with a luciferase based system

被引:18
|
作者
Allegra, Danilo
Mertens, Daniel [1 ]
机构
[1] Univ Hosp, Dept Internal Med 3, D-89081 Ulm, Germany
关键词
MicroRNA; Luciferase; miRNA processing assay; Drosha; Microprocessor; miR-16; RNA; MATURATION; RECOGNITION; COMPLEX; REGION; SUBSET; DICER;
D O I
10.1016/j.bbrc.2011.02.055
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The RNAse III Drosha is responsible for the first step of microRNA maturation, the cleavage of primary miRNA to produce the precursor miRNA. Processing by Drosha is finely regulated and influences the amount of mature microRNA in a cell. We describe in the present work a method to quantify Drosha processing activity in-vivo, which is applicable to any microRNA. With respect to other methods for measuring Drosha activity, our system is faster and scalable, can be used with any cellular system and does not require cell sorting or use of radioactive isotopes. This system is useful to study regulation of Drosha activity in physiological and pathological conditions. (C) 2011 Elsevier Inc. All rights reserved.
引用
收藏
页码:501 / 505
页数:5
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