Transcriptomic profiling of circadian genes in WT mouse tissues (mRNA-Seq)

A goal of a collaboration between the CHDI Foundation and Prof. Jenny Morton at Cambridge University was to identify genes in mice with diurnal/circadian expression patterns. Tissues from wild type mice were harvested every 3 hrs over a 36 hour time period, for a total of 13 time points. Striatal, cortical, hypothalamic and liver samples were harvested, flash frozen, and transferred to Expression Analysis for RNA extraction and mRNA-seq sequencing. Samples passing QC were analyzed to produce high quality, 50-base paired-end RNA-seq results. Striatal, cortical, hypothalamic and liver samples were collected from 26-week-old wild type C57BL/6J male mice every 3 hours for 36 hours, number of replicates 2 to 12 for each ZT group. mRNA-Seq was performed.CHDI基金会与剑桥大学珍妮·莫顿（Jenny Morton）教授的合作项目，核心目标之一是鉴定出具有昼行/昼夜节律（diurnal/circadian）表达模式的小鼠基因。研究团队在36小时的周期内，每3小时采集一次野生型小鼠的组织样本，共计13个时间点。本次采集的组织涵盖纹状体、皮层、下丘脑与肝脏样本，经快速冷冻后送至专业表达分析机构进行RNA提取与mRNA-seq测序。通过质量控制（Quality Control, QC）的样本经测序分析，最终获得了高质量的50碱基双端RNA-seq测序结果。本研究的采样对象为26周龄的野生型C57BL/6J雄性小鼠，同样以每3小时一次的频率采集上述四类组织，持续36小时；每个授时因子时间（Zeitgeber Time, ZT）组的生物学重复数为2至12次，最终完成了mRNA-seq测序。