m6A/m-Seq of mouse adult cortex of Mettl3 cKO or Fto cKO mice

N6-Methyladenosine (m6A) and N6,2'-O-dimethyladenosine (m6Am) are abundant mRNA modifications that regulate transcript processing and translation. The role of both, here termed m6A/m, in the stress response in the adult brain in vivo are currently unknown. Here, we investigated the effect of gene deletion of Mettl3, a m6A methyltransferase, and Fto, a m6A and m6Am demethlyase, induced in adulthood in excitatory neurons of the neocortex and hippocampus (Camk2a-Cre Mettl3 or Fto cKO) on the cortical epitranscriptome. PolyA-RNA-fragments from 3-5 replicates per group were processed both as m6A/m-sample (RNA immunoprecipiation RIP with an m6A and m6Am antibody) and RNA-input sample. 5 replicates of WT mice (from both lines) as well as 3 replicates of Mettl3 cKO and 3 replicates of Fto cKO mice (cortex sample from 3 mice pooled per replicate) were processed as m6A/m-immunoprecipitation-sample ("m6A") or as input RNA samples ("RNA"). Additionally, 1 RNA sample (equimolar mixed from all samples) was processed as IgG control.N6-甲基腺苷（N6-Methyladenosine，m6A）与N6,2'-O-二甲基腺苷（N6,2'-O-dimethyladenosine，m6Am）是两类丰富的mRNA修饰方式，可调控转录本加工与翻译过程。目前，本文中将二者统称为m6A/m的这两类修饰在成年大脑体内应激反应中的作用尚不明确。本研究针对成年小鼠新皮层与海马体兴奋性神经元中诱导性条件性敲除（Camk2a-Cre介导的Mettl3或Fto cKO）的两个基因——m6A甲基转移酶Mettl3，以及m6A和m6Am去甲基化酶Fto——展开研究，探究其对皮层表观转录组的影响。每组取3~5个生物学重复的PolyA-RNA片段，分别以m6A/m样本（采用针对m6A与m6Am的抗体进行RNA免疫沉淀（RIP））和RNA输入样本进行处理。本研究同时对5个野生型（wild type，WT）小鼠重复样本（来自两种品系）、3个Mettl3条件性敲除（Mettl3 cKO）小鼠重复样本，以及3个Fto条件性敲除（Fto cKO）小鼠重复样本（每个重复样本由3只小鼠的皮层组织混合制备）进行处理，分别作为m6A/m免疫沉淀样本（标记为"m6A"）或输入RNA样本（标记为"RNA"）。此外，我们将1份由所有样本等摩尔混合制备的RNA样本作为IgG对照样本进行处理。