The effects of METTL3 on BAT development and thermogenesis

Purpose: The goal of this study is to compare transcriptome profilings of interscapular brown adipose tissue (iBAT) from BAT-specific METTL3 knockout and control mice Methods: iBATs were collected from METTL3 BKO and f/f mice at 8 weeks old, and total RNA was isolated using TriPure. RNA samples were pooled from iBATs of METTL3 BKO and f/f mice (n=4 for each group). RNA-seq was performed by using Illumina NovaSeq 6000 platform. Paired-end clean reads were aligned to the mouse reference genome (Ensemble_GRCm38.90) 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 METTL3 BKO and f/f mice, generated by RNA-seq technology. The RNA-seq analysis showed that 530 genes were down-regulated and 1552 genes were up-regulated in the iBAT of METTL3 BKO mice. GO analysis showed that the down-regulated genes were primarily related to developmental maturation, respiratory electron transport chain, adaptive thermogenesis, and energy deprivation by oxidation of organic compounds whereas the up-regulated genes were associated with muscle system process and muscle cell development. Overall design: RNA samples were pooled from iBATs of METTL3 BKO and f/f mice at 8 weeks old (n=4 for each group). mRNA profiles of METTL3 BKO and f/f iBATs were generated by deep sequencing, in a single test, using Illumina NovaSeq 6000 platform.