The effects of ALKBH5 on liver function [RNA-seq]

The goal of this study is to investigate how ALKBH5 regulates liver function. mRNA profiles in livers of Alkbh5flox/flox and Alkbh5-HKO mice were generated by deep sequencing using Illumina Novaseq 6000 platform (n=3 for each group). Methods: Total RNA was extracted using Tripure Isolation Reagent (Roche, Mannheim, Germany) from livers of Alkbh5flox/flox and Alkbh5-HKO mice at 9 weeks old. 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.p6) 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 liver transcriptomes from Alkbh5flox/flox and Alkbh5-HKO mice, generated by RNA-seq technology. The RNA-seq analysis showed that 1107 genes were downregulated and 1248 genes were upregulated in the livers of Alkbh5-HKO mice.

本研究旨在探究ALKBH5对肝功能的调控机制。研究通过Illumina Novaseq 6000测序平台开展深度测序，获取了Alkbh5flox/flox与Alkbh5-HKO小鼠肝脏组织的信使RNA（mRNA）表达谱（每组n=3）；实验方法如下：于9周龄时，采用Tripure Isolation Reagent（罗氏，德国曼海姆）从两组小鼠肝脏中提取总RNA，每组设置3个独立生物学重复，随后使用Illumina Novaseq 6000测序平台完成RNA测序（RNA-seq），将配对端清洁读段（Paired-end clean reads）比对至小鼠参考基因组GRCm38.p6（比对工具为Hisat2 v2.0.5），再通过featureCounts v1.5.0-p3对比对得到的读段进行mRNA表达量定量。本研究为首次利用RNA测序（RNA-seq）技术对Alkbh5flox/flox与Alkbh5-HKO小鼠肝脏转录组开展的详细分析，RNA测序分析结果显示，Alkbh5-HKO小鼠肝脏组织中存在1107个下调基因与1248个上调基因。