植物 miRNA 初级转录本的定量分析。

Quantitative Analysis of Plant miRNA Primary Transcripts.

机构信息

Department of Gene Expression, Institute of Molecular Biology and Biotechnology, Faculty of Biology, Adam Mickiewicz University, Poznań, Poznań, Poland.

Department of Computational Biology, Institute of Molecular Biology and Biotechnology, Faculty of Biology, Adam Mickiewicz University, Poznań, Poznań, Poland.

出版信息

Methods Mol Biol. 2021;2170:53-77. doi: 10.1007/978-1-0716-0743-5_5.

DOI:10.1007/978-1-0716-0743-5_5

PMID:32797451

Abstract

MicroRNAs control plant development and are key regulators of plant responses to biotic and abiotic stresses. Thus, their expression must be carefully controlled since both excess and deficiency of a given microRNA may be deleterious to plant cell. MicroRNA expression regulation can occur at several stages of their biogenesis pathway. One of the most important of these regulatory checkpoints is transcription efficiency. mirEX database is a tool for exploration and visualization of plant pri-miRNA expression profiles. It includes results obtained using high-throughput RT-qPCR platform designed to monitor pri-miRNA expression in different miRNA biogenesis mutants and developmental stages of Arabidopsis, barley, and Pellia plants. A step-by-step instruction for browsing the database and detailed protocol for high-throughput RT-qPCR experiments, including list of primers designed for the amplification of pri-miRNAs, are presented.

摘要

MicroRNAs 控制植物的发育，是植物对生物和非生物胁迫反应的关键调节剂。因此，它们的表达必须被仔细控制，因为给定 microRNA 的过量和缺乏都可能对植物细胞有害。microRNA 的表达调控可以发生在它们生物发生途径的几个阶段。其中最重要的调控检查点之一是转录效率。mirEX 数据库是一个探索和可视化植物 pri-miRNA 表达谱的工具。它包括使用高通量 RT-qPCR 平台获得的结果，该平台旨在监测拟南芥、大麦和 Pellia 植物不同 miRNA 生物发生突变体和发育阶段的 pri-miRNA 表达。提供了浏览数据库的分步说明和高通量 RT-qPCR 实验的详细方案，包括用于 pri-miRNAs 扩增的引物列表。