m6A mRNA modification is a rheostat to maintain colonic epithelial cell homeostasis [bulkRNA-seq]

The function of m6A modification in colonic epithelial cells has been validated in our study that depletion of METTL14 in colonic epithelial cells triggers cell death. To further explore the biological effects of m6A deficiency in colonic epithelial cells, we performed RNA sequencing analysis of colonic epithelial cells form 2-week-old Mettl14-KO and WT control mice.The bioinformatic analysis revealed that METTL14 depletion increased the mRNA expression of NF-kB inhibitor Nfkbia, which favored TNF-induced cell death. RNA samples from Mettl14-KO and WT control colonic epithelial cells were collected, and then RNA purification, reverse transcription, library construction and sequencing were performed according to the manufacturer's instructions (Illumina).本研究证实了m6A修饰在结肠上皮细胞中的生物学功能：结肠上皮细胞中METTL14（甲基转移酶样蛋白14）的缺失会触发细胞死亡。为进一步探究结肠上皮细胞中m6A缺失的生物学效应，我们对2周龄Mettl14-KO小鼠与野生型（WT）对照小鼠的结肠上皮细胞开展了RNA测序分析。生物信息学分析显示，METTL14缺失可上调核因子κB（NF-κB）抑制剂Nfkbia的mRNA表达水平，该变化会增强肿瘤坏死因子（TNF）诱导的细胞死亡。我们收集了Mettl14-KO与WT对照小鼠的结肠上皮细胞RNA样本，并按照制造商Illumina的操作指南完成了RNA纯化、逆转录、文库构建及测序工作。