BRIC-seq：一种用于确定哺乳动物细胞中RNA稳定性的全基因组方法。

BRIC-seq: a genome-wide approach for determining RNA stability in mammalian cells.

作者信息

Imamachi Naoto, Tani Hidenori, Mizutani Rena, Imamura Katsutoshi, Irie Takuma, Suzuki Yutaka, Akimitsu Nobuyoshi

机构信息

Radioisotope Center, The University of Tokyo, 2-11-16 Yayoi, Bunkyo-ku, Tokyo 113-0032, Japan.

Research Institute for Environmental Management Technology, National Institute of Advanced Industrial Science and Technology (AIST), 16-1 Onogawa, Tsukuba 305-8569, Japan.

出版信息

Methods. 2014 May 1;67(1):55-63. doi: 10.1016/j.ymeth.2013.07.014. Epub 2013 Jul 17.

DOI:10.1016/j.ymeth.2013.07.014

PMID:23872059

Abstract

We recently developed a novel transcriptome analysis method, termed 5'-bromo-uridine (BrU) immunoprecipitation chase-deep sequencing analysis (BRIC-seq). BRIC-seq enables the determination of genome-wide RNA stability by chasing chronological decreases of BrU-labeled RNAs under physiologically undisturbed conditions. The RNA half-life of each transcript is calculated from the decreasing number of BrU-labeled RNA sequence tags measured by deep sequencing of BrU-labeled RNAs. Here, we describe a detailed protocol and provide tips for BRIC-seq, followed by computational analysis.

摘要

我们最近开发了一种新型转录组分析方法，称为5'-溴尿苷（BrU）免疫沉淀追踪深度测序分析（BRIC-seq）。BRIC-seq能够在生理条件未受干扰的情况下，通过追踪BrU标记RNA随时间的减少来测定全基因组范围内的RNA稳定性。每个转录本的RNA半衰期是根据对BrU标记RNA进行深度测序所测得的BrU标记RNA序列标签数量的减少来计算的。在此，我们描述了BRIC-seq的详细方案并提供了相关提示，随后是计算分析。