Drosha复合物活性的体外和体内测定

In vitro and in vivo assays for the activity of Drosha complex.

作者信息

Lee Yoontae, Kim V Narry

机构信息

School of Biological Sciences, Seoul National University, Seoul, Korea.

出版信息

Methods Enzymol. 2007;427:89-106. doi: 10.1016/S0076-6879(07)27005-3.

DOI:10.1016/S0076-6879(07)27005-3

PMID:17720480

Abstract

MicroRNA (miRNA) genes are transcribed into long primary transcripts (pri-miRNAs) that get processed into mature miRNAs of about 22 nt in length by two different ribonuclease (RNase) III enzymes, Drosha and Dicer. Various experimental protocols have been developed and modified for genetic and biochemical analyses for microRNA processing. Here we describe the methods for the analysis of pri-miRNA processing that is mediated by Drosha and its cofactor, DiGeorge Syndrome Critical Region Gene 8 (DGCR8).

摘要

微小RNA（miRNA）基因转录生成长长的初级转录本（pri-miRNA），这些转录本通过两种不同的核糖核酸酶（RNase）III 酶—— Drosha和Dicer，被加工成长度约为22个核苷酸的成熟miRNA。已经开发并修改了各种实验方案，用于对微小RNA加工进行遗传和生化分析。在这里，我们描述了分析由Drosha及其辅因子——22q11.2微缺失综合征关键区域基因8（DGCR8）介导的pri-miRNA加工的方法。