Dysregulated ac4C modification of mRNA in a mouse model of early stage Alzheimer's disease [acRIP-seq]

In this study, ac4C modifications in mRNA were investigated using three-month-old 5×FAD transgenic mice, a widely recognized model of Alzheimer's disease, along with age- and sex-matched wild-type (WT) controls. To elucidate the role of ac4C-modified mRNA in AD, we employed three analytical techniques: acetylated RNA immunoprecipitation sequencing (ac4C-RIP-seq), RNA sequencing (RNA-seq), and proteomic analysis. The first two were used to identify mRNAs carrying ac4C modifications and to quantify mRNA abundance, respectively. Overall design: acetylated RNA immunoprecipitation -sequancing (acRIP-seq) was used to determine the acetylation modification of mRNA in the hippocampus from 3 month mice (3 5xFAD mice and 3 WT mice). Then IgG- sequancing was used to correct the result of acRIP-seq.

本研究以3月龄5xFAD转基因小鼠——该模型为当前广泛认可的阿尔茨海默病（Alzheimer's disease, AD）经典模型——及年龄、性别匹配的野生型（wild-type, WT）对照小鼠为研究对象，系统探究了mRNA上的ac4C修饰（ac4C modification）。为明确ac4C修饰的mRNA在阿尔茨海默病进程中的功能，本研究采用了三类分析技术：乙酰化RNA免疫沉淀测序（acetylated RNA immunoprecipitation sequencing, ac4C-RIP-seq）、RNA测序（RNA sequencing, RNA-seq）与蛋白质组学分析。其中前两种技术分别用于筛选携带ac4C修饰的mRNA分子，以及定量检测其表达丰度。
整体实验设计如下：首先利用乙酰化RNA免疫沉淀测序（acRIP-seq），对3月龄小鼠（3只5xFAD小鼠与3只WT小鼠）的海马体组织中mRNA的乙酰化修饰水平进行检测；随后通过IgG测序（IgG sequencing）对acRIP-seq的实验结果进行背景校正。