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. 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：本研究旨在利用细菌人工染色体-核糖体亲和纯化技术（BAC-TRAP）以及携带酪氨酸羟化酶（Th）启动子驱动Cre重组酶的腺相关病毒（AAV:THCre），解析小鼠中脑多巴胺能神经元内雌激素相关受体γ（Esrrg）基因敲除相关的基因表达变化。
实验方法：所有受试小鼠均携带用于BAC-TRAP RNA分离的L10a转基因，通过AAV:THCre介导实现Esrrg基因敲除或不进行基因敲除。待小鼠注射病毒后饲养1个月，摘取其中脑组织并置于干冰上快速冷冻。采用BAC-TRAP法分离RNA后，将样本送往测序平台进行测序。本实验对中脑Esrrg基因敲除小鼠与未敲除小鼠的中脑mRNA表达谱进行比较分析，所有受试小鼠均携带L10a转基因以适配BAC-TRAP RNA分离流程。