Calarco John A, Taylor Seth R, Miller David M
Department of Cell and Systems Biology, University of Toronto, Toronto, ON, Canada, M5S 3G5.
Department of Cell Biology and Physiology, Brigham Young University, Provo, UT 84602, USA.
Genetics. 2025 Jan 8;229(1):1-108. doi: 10.1093/genetics/iyae167.
Reliable methods for detecting and analyzing gene expression are necessary tools for understanding development and investigating biological responses to genetic and environmental perturbation. With its fully sequenced genome, invariant cell lineage, transparent body, wiring diagram, detailed anatomy, and wide array of genetic tools, Caenorhabditis elegans is an exceptionally useful model organism for linking gene expression to cellular phenotypes. The development of new techniques in recent years has greatly expanded our ability to detect gene expression at high resolution. Here, we provide an overview of gene expression methods for C. elegans, including techniques for detecting transcripts and proteins in situ, bulk RNA sequencing of whole worms and specific tissues and cells, single-cell RNA sequencing, and high-throughput proteomics. We discuss important considerations for choosing among these techniques and provide an overview of publicly available online resources for gene expression data.
可靠的基因表达检测与分析方法是理解发育过程以及研究生物体对遗传和环境扰动的生物学反应的必要工具。秀丽隐杆线虫具有全基因组测序、恒定的细胞谱系、透明的身体、连接图谱、详细的解剖结构以及丰富多样的遗传工具,是将基因表达与细胞表型联系起来的极为有用的模式生物。近年来新技术的发展极大地扩展了我们以高分辨率检测基因表达的能力。在此,我们概述了秀丽隐杆线虫的基因表达方法,包括原位检测转录本和蛋白质的技术、全虫及特定组织和细胞的批量RNA测序、单细胞RNA测序以及高通量蛋白质组学。我们讨论了在这些技术中进行选择时的重要考量因素,并概述了可公开获取的基因表达数据在线资源。