Lau Nelson C
Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts, USA.
Curr Protoc Mol Biol. 2008 Jan;Chapter 26:Unit26.7. doi: 10.1002/0471142727.mb2607s81.
Eukaryotic cells express small noncoding RNAs to silence target genes or regulate chromatin domains. MicroRNAs (miRNA) are endogenous small RNAs that are thought to each regulate multiple mRNA targets. To gain a deeper understanding of processes regulated by small noncoding RNAs, techniques are required to identify and detect them. This unit describes standard laboratory methods for and comments about high-throughput technologies for the identification and detection of small RNAs. This unit also outlines a cell-based reporter gene assay for assessing the regulatory potential of a microRNA on a candidate mRNA target.
真核细胞表达小的非编码RNA以沉默靶基因或调控染色质结构域。微小RNA（miRNA）是内源性小RNA，据认为每个miRNA可调控多个mRNA靶标。为了更深入地了解由小非编码RNA调控的过程，需要用于鉴定和检测它们的技术。本单元描述了用于鉴定和检测小RNA的高通量技术的标准实验室方法及相关评论。本单元还概述了一种基于细胞的报告基因检测方法，用于评估微小RNA对候选mRNA靶标的调控潜力。
