Institute for Research in Biomedicine (IRB Barcelona), The Barcelona Institute of Science and Technology (BIST), 08028 Barcelona, Spain.
Hospital del Mar Medical Research Institute (IMIM), Cancer Research Program, 08003 Barcelona, Spain.
STAR Protoc. 2021 May 14;2(2):100539. doi: 10.1016/j.xpro.2021.100539. eCollection 2021 Jun 18.
Molecular daily rhythms can be captured by precisely timed tissue harvests from groups of animals. This protocol will allow the investigator to identify transcriptional rhythms in the mouse liver while also providing a template for similar analyses in other whole metabolic organs. We describe steps for mouse entrainment, liver dissection, and rhythmicity analysis from total RNA sequencing data. The resulting rhythmic transcriptome will provide the user with a starting point for defining specific biological processes that undergo daily rhythms. For complete details on the use and execution of this protocol, please refer to Koronowski et al. (2019). A similar protocol for interfollicular epidermal cells is demonstrated in Welz et al. (2019).
分子的日常节律可以通过从动物群体中精确计时采集组织来捕捉。该方案将允许研究人员在鉴定小鼠肝脏的转录节律的同时,为其他整个代谢器官的类似分析提供模板。我们描述了从小鼠肝脏总 RNA 测序数据中进行节律性分析的小鼠驯化、肝脏解剖和分析步骤。由此产生的节律转录组将为用户提供一个起点,用于定义经历日常节律的特定生物学过程。有关本方案使用和执行的完整详细信息,请参考 Koronowski 等人(2019 年)。Welz 等人(2019 年)展示了类似的滤泡间表皮细胞方案。