小RNA在体内对靶向mRNA稳定性的作用。

Activity of small RNAs on the stability of targeted mRNAs in vivo.

作者信息

Desnoyers Guillaume, Massé Eric

机构信息

Department of Biochemistry, University of Sherbrooke, Sherbrooke, QC, Canada.

出版信息

Methods Mol Biol. 2012;905:245-55. doi: 10.1007/978-1-61779-949-5_16.

DOI:10.1007/978-1-61779-949-5_16

PMID:22736009

Abstract

Northern blots are extremely useful to monitor the steady state level of small regulatory RNAs (sRNAs) as well as their target mRNAs. In combination with the drug rifampicin, which blocks cellular transcription, Northern blots can be used to determine the stability of sRNAs and mRNAs. Here we describe a protocol to assess the activity of the sRNA RyhB on the stability of targeted mRNAs sodB, fumA, and iscRSUA. We also describe how to identify a sRNA-induced initial cleavage site on a target mRNA. This protocol can be used for any sRNAs and their target mRNAs.

摘要

Northern印迹对于监测小调节RNA（sRNA）及其靶标mRNA的稳态水平极为有用。结合可阻断细胞转录的利福平药物，Northern印迹可用于确定sRNA和mRNA的稳定性。在此，我们描述了一种评估sRNA RyhB对靶标mRNA sodB、fumA和iscRSUA稳定性的活性的方法。我们还描述了如何鉴定靶标mRNA上sRNA诱导的初始切割位点。该方法可用于任何sRNA及其靶标mRNA。