High-resolution transcriptome mapping and N6-methyladenosine modification mapping of 3T3-L1 derived adipocytes (RNA-Seq). High-resolution transcriptome mapping and N6-methyladenosine modification mapping of 3T3-L1 derived adipocytes (RNA-Seq)

Purpose:Detect transcriptome-wide mapping of m6A modifications by MeRIP-seq between Rhein and DMSO treated 3T3-L1 differentiation adipocytes at the preadipocytes, MCE stage and differentiation stage. Methods:Total RNA was extracted using TRIzol Reagent followed by chemical fragmentation. The fragmented RNA was incubated with anti-m6A antibody. The desired m6A-enriched RNA eluate was used to construct cDNA library in parallel with input control. Conclusion: Our study revealed the difference of m6A-methylated RNA between DMSO and Rhein treated cells by RNA-seq and MeRIP-seq technologies. Overall design: Compare the difference of m6A-methyalated RNA at the preadipocytes, MCE stage and differentiation stage during differentiation between DMSO and Rhein treated samples.

研究目的：采用甲基化RNA免疫沉淀测序（MeRIP-seq）技术，针对大黄酸（Rhein）处理组与二甲基亚砜（Dimethyl Sulfoxide，DMSO）处理组的3T3-L1分化脂肪细胞，在前脂肪细胞期、有丝分裂克隆扩增（MCE）阶段及分化阶段开展全转录组范围的N6-甲基腺苷（m6A）修饰定位检测。
实验方法：使用TRIzol试剂提取总RNA，随后进行化学片段化处理；将片段化后的RNA与抗m6A抗体孵育，收集目标m6A富集RNA洗脱液，并与输入对照（input control）并行构建cDNA文库。
研究结论：本研究通过RNA测序（RNA-seq）与MeRIP-seq技术，揭示了DMSO与Rhein处理的细胞间m6A甲基化RNA的修饰差异。
整体实验设计：对比脂肪细胞分化过程中，DMSO处理组与Rhein处理组的样本在前脂肪细胞期、MCE阶段及分化阶段的m6A甲基化RNA差异。