The effects of WTAP on heart development [WTAPCKO]. The effects of WTAP on heart development [WTAPCKO]

The goal of this study is to investigate how WTAP regulates heart development. Overall design: mRNA profiles in the hearts of Wtapflox/flox and Wtap-CKO mice were generated by deep sequencing using Illumina Novaseq 6000 platform (n=3 for each group). Total RNA was extracted using Tripure Isolation Reagent (Roche, Mannheim, Germany) from the hearts of Wtapflox/flox and Wtap-CKO mice at 4 days 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(GRCm39) 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 heart transcriptomes from Wtapflox/flox and Wtap-CKO mice, generated by RNA-seq technology. The RNA-seq analysis showed that 2348 genes were downregulated and 2764 genes were upregulated in the hearts of Wtap-CKO mice.

本研究旨在探究WTAP对心脏发育的调控机制。总体实验设计如下：通过Illumina Novaseq 6000平台开展深度测序，获取Wtapflox/flox及Wtap-CKO小鼠心脏组织的mRNA表达谱（每组设置3个生物学重复）。实验材料为4日龄Wtapflox/flox与Wtap-CKO小鼠的心脏组织，采用Tripure Isolation Reagent（罗氏，德国曼海姆）完成总RNA提取。每组设置3个独立生物学重复用于RNA测序，本次RNA测序采用Illumina Novaseq 6000平台完成深度测序。采用Hisat2 v2.0.5软件将双端清洁reads比对至小鼠参考基因组GRCm39，随后通过featureCounts v1.5.0-p3软件对比对得到的reads进行mRNA表达量定量。本研究为首次通过RNA测序技术对Wtapflox/flox及Wtap-CKO小鼠的心脏转录组开展的系统性详细分析。RNA测序分析结果显示，在Wtap-CKO小鼠的心脏组织中，共有2348个基因表达下调，2764个基因表达上调。