Gene changes associated with deletion of Esrrg in Dopaminergic neurons in mice [H202SC21050050]. Gene changes associated with deletion of Esrrg in Dopaminergic neurons in mice [H202SC21050050]

Purpose: The goal of this study is to understand what gene changes are associated with removal of Esrrg in dopaminerigc neurons from the mouse midbrain using BAC-TRAP and an AAV for Thcre Methods: Midbrain from mice expressing the L10a transgene used for BAC-TRAP method of RNA isolation both with and without deletion of Esrrg using AAV:THCre were aged 1 month post-injection and midbrains were then extracted and flash frozen on dry ice. RNA was isolated using the BAC-TRAP method and samples were sent for sequencing. Overall design: Midbrain mRNA profiles of mice 1 month post deletion of Esrrg in the midbrain compared to mice with no Esrrg deletion. All mice were expressing the L10a transgene to allow for BAC-TRAP method of RNA-isolation.

研究目的：本研究旨在利用BAC-TRAP技术与靶向酪氨酸羟化酶启动子的Cre重组酶腺相关病毒（AAV:TH-Cre），解析小鼠中脑多巴胺能神经元中Esrrg基因敲除相关的基因表达变化。
实验方法：实验对象为携带L10a转基因的小鼠，该转基因可用于BAC-TRAP法进行RNA分离。通过AAV:TH-Cre对小鼠中脑多巴胺能神经元进行Esrrg基因敲除，设置未进行基因敲除的对照组；所有小鼠在病毒注射后饲养1个月，随后提取中脑组织并置于干冰上快速冷冻。采用BAC-TRAP法分离总RNA，将样本送检进行高通量测序。
实验设计：本实验对比两组小鼠的中脑mRNA表达谱：一组为中脑Esrrg基因敲除组（病毒注射后1个月），另一组为未进行Esrrg基因敲除的对照组。所有实验小鼠均携带L10a转基因，以支持BAC-TRAP法完成RNA分离。