Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA, USA.
Harvard Stem Cell Institute, Cambridge, MA, USA.
Methods Mol Biol. 2023;2693:39-60. doi: 10.1007/978-1-0716-3342-7_4.
RNA sequencing (RNA-seq) is a powerful method of transcriptional analysis that allows for the sequence identification and quantification of cellular transcripts. RNA-seq can be used for differential gene expression (DGE) analysis, gene fusion detection, allele-specific expression, isoform and splice variant quantification, and identification of novel genes. These applications can be used for downstream systems biology analyses such as gene ontology or pathway analysis to provide insight into processes altered between biological conditions. Given the wide range of signaling pathways subject to chaperone activity as well as numerous chaperone functions in RNA metabolism, RNA-seq may provide a valuable tool for the study of chaperone proteins in biology and disease. This chapter outlines an example RNA-seq workflow to determine differentially expressed (DE) genes between two or more sample conditions and provides some considerations for RNA-seq experimental design.
RNA 测序（RNA-seq）是一种强大的转录分析方法，可用于鉴定和定量细胞转录本的序列。RNA-seq 可用于差异基因表达（DGE）分析、基因融合检测、等位基因特异性表达、异构体和剪接变体定量以及新基因的鉴定。这些应用可用于下游系统生物学分析，如基因本体论或途径分析，以深入了解生物条件之间改变的过程。鉴于广泛的信号通路受到伴侣活性的影响，以及 RNA 代谢中的许多伴侣功能，RNA-seq 可能为研究生物学和疾病中的伴侣蛋白提供了有价值的工具。本章概述了一个示例 RNA-seq 工作流程，用于确定两个或更多样本条件之间差异表达（DE）的基因，并提供了一些关于 RNA-seq 实验设计的考虑因素。
