The effects of WTAP on islet β-cell function

The goal of this study is to investigate how WTAP regulates islet β-cell function. Islets were isolated from pancreatic islets of Wtapflox/flox and Wtap-βKO mice at 7 weeks old. One islet sample was combined from three mice. Total RNA was extracted using Tripure Isolation Reagent (Roche, Mannheim, Germany). Three independent biological replicates for each group were used for RNA-seq. RNA-seq was performed by deep sequencing using an Illumina Novaseq 6000 platform. Paired-end clean reads were aligned to the mouse reference genome(GRCm38/mm10) with Hisat2 v2.0.5, and the aligned reads were used to quantify mRNA expression by using featureCounts v1.5.0-p3. Our study represents the first detailed analysis of islet transcriptomes from Wtapflox/flox and Wtap-βKO mice, generated by RNA-seq technology. The RNA-seq analysis showed that 3015 genes were downregulated and 2900 genes were upregulated in the pancreatic islets of Wtap-βKO mice. mRNA profiles in pancreatic islets of Wtapflox/flox and Wtap-βKO mice were generated by deep sequencing using Illumina Novaseq 6000 platform (n=3 for each group).

本研究旨在探究WTAP对胰岛β细胞功能的调控机制。我们从7周龄的Wtapflox/flox与Wtap-βKO小鼠的胰腺胰岛中分离得到胰岛组织，每份样本均由3只小鼠的胰岛组织混合制备。采用德国曼海姆罗氏（Roche）公司的Tripure Isolation Reagent提取总RNA。每组设置3次独立生物学重复用于RNA测序（RNA-seq）实验，测序工作依托Illumina Novaseq 6000平台开展深度测序。使用Hisat2 v2.0.5软件将双端clean reads比对至小鼠参考基因组（GRCm38/mm10），并通过featureCounts v1.5.0-p3工具对mRNA表达水平进行定量分析。本研究为首项借助RNA测序技术，针对Wtapflox/flox与Wtap-βKO小鼠胰岛转录组开展的详细分析工作。RNA测序分析结果显示，Wtap-βKO小鼠胰腺胰岛中共有3015个基因表达下调、2900个基因表达上调。本研究通过Illumina Novaseq 6000平台完成了Wtapflox/flox与Wtap-βKO小鼠胰腺胰岛的mRNA表达谱测序，每组样本量n=3。