The effects of WTAP on BAT function

Purpose: The goal of this study is to compare transcriptome profilings of interscapular brown adipose tissue (iBAT) from BAT-specific WTAP knockout and control mice Methods: iBATs were collected from WTAP BKO and f/f mice at 8 weeks old (n=3 for each group), and total RNA was isolated using TriPure. RNA-seq was performed by using Illumina NovaSeq 6000 platform. Paired-end clean reads were aligned to the mouse reference genome (Ensemble_grcm38_p6) with TopHat (version 2.0.12), and the aligned reads were used to quantify mRNA expression by using HTSeq-count (version 0.6.1). Conclusion:Our study represents the first detailed analysis of iBAT transcriptomes from WTAP BKO and f/f mice, generated by RNA-seq technology. The RNA-seq analysis showed that 894 genes were down-regulated and 1410 genes were up-regulated in the iBAT of WTAP BKO mice. GO analysis showed that the down-regulated genes were primarily related to related to generation of precursor metabolites and energy, cellular respiration, energy derivation by oxidation of organic compounds, electron transport chain, nucleotide metabolic process and purine ribonucleotide metabolic process. RNA samples were isolated from iBATs of WTAP BKO and f/f mice at 8 weeks old (n=3 for each group). mRNA profiles of WTAP BKO and f/f iBATs were generated by deep sequencing using Illumina NovaSeq 6000 platform.研究目的：本研究旨在对比BAT特异性WTAP敲除小鼠与对照小鼠肩胛间棕色脂肪组织（interscapular brown adipose tissue，iBAT）的转录组谱。 实验方法：于8周龄时从WTAP BKO小鼠及f/f对照小鼠中采集肩胛间棕色脂肪组织（iBAT），每组设置3个生物学重复（n=3）；采用TriPure试剂提取总RNA。使用Illumina NovaSeq 6000测序平台开展RNA测序。将双端洁净reads比对至小鼠参考基因组（Ensemble_grcm38_p6），比对工具为TopHat（版本2.0.12）；随后利用HTSeq-count（版本0.6.1）对比对得到的reads进行mRNA表达量定量。 研究结论：本研究首次通过RNA测序技术，对WTAP BKO及f/f小鼠的肩胛间棕色脂肪组织转录组进行了详细分析。RNA测序结果显示，WTAP BKO小鼠的iBAT中共有894个基因表达下调、1410个基因表达上调。基因本体（Gene Ontology，GO）分析结果表明，下调表达的基因主要与前体代谢物与能量生成、细胞呼吸、有机化合物氧化供能、电子传递链、核苷酸代谢过程以及嘌呤核糖核苷酸代谢过程相关。 于8周龄时从WTAP BKO及f/f小鼠中采集肩胛间棕色脂肪组织并提取总RNA样本，每组设置3个生物学重复（n=3）。利用Illumina NovaSeq 6000测序平台开展深度测序，获得了WTAP BKO与f/f小鼠iBAT的mRNA表达谱。