Department of Molecular Biology, Princeton University, Princeton, NJ, USA.
Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, NJ, USA.
Methods Mol Biol. 2020;2144:177-186. doi: 10.1007/978-1-0716-0592-9_16.
Multicellular organisms are composed of distinct cells and tissues that coordinate highly orchestrated responses to environmental challenges, including those that arise with age. Since C. elegans is a premier model system used to study the molecular and cellular regulators of adult and aging phenotypes, cell type- and tissue-specific approaches are needed to characterize the genome-wide expression changes associated with these responses. Here we describe a method for the FACS-based isolation and RNA sequencing of dissociated cells from adult C. elegans. This technique is amenable to profiling the cell- and tissue-specific gene expression changes in C. elegans mutants, including aging models, such as the daf-2/insulin-like signaling (IIS) pathway, and in wild-type animals with age.
多细胞生物由不同的细胞和组织组成，这些细胞和组织协调对环境挑战的高度协调反应，包括随着年龄增长而出现的挑战。由于秀丽隐杆线虫是用于研究成年和衰老表型的分子和细胞调节剂的主要模型系统，因此需要采用细胞类型和组织特异性方法来描述与这些反应相关的全基因组表达变化。在这里，我们描述了一种基于流式细胞术从成年秀丽隐杆线虫中分离和 RNA 测序的方法。该技术适用于分析秀丽隐杆线虫突变体（包括衰老模型，如 daf-2/胰岛素样信号传导 (IIS) 途径）以及随着年龄增长的野生型动物的细胞和组织特异性基因表达变化。
