Next Generation Sequencing Facilitates Quantitative Analysis of Germ Cell-specific Ythdf2 Mutants and Control Transcriptomes

Given the reported molecular role of YTHDF2 in mRNA stability, we designed to investigate the biological mechanism underlying the defects of germ cell-specific ythdf2 mutants (Ythdf2-vKO) spermatogenesis using high-throughput RNA sequencing (RNA-seq). Testes were obtained from Ythdf2-vKO and control adult mice, then were used for RNA-seq. Our result showed that there were 266 genes (196 up and 70 down) deregulated in Ythdf2-vKO testes (P < 0.05, fold change ＞ 1.5) compared with control adult mice. Based on our previously reported m6A mRNA methylomes of mouse spermaogenic cells (PMID: 28914256), 66% of the upregulated genes (129 genes) in Ythdf2-vKO testes were modified with m6A. Further analysis based on our previously reported single-cell transcriptomics data of mouse spermatogenesis (PMID: 30061742), the 129 upregulated genes with m6A modification in Ythdf2-vKO testes mainly express at early stages of wild-type spermatogenesis. While the 70 down regulated genes in Ythdf2-vKO testes mainly exist at later stages of wild-type spermatogenesis. Our qPCR analysis confirmed the RNA-seq analysis. In summary, our results showed that YTHDF2 is essential for regulation of a wave of transcriptome dynamic transition during mice spermatogenesis. Transcriptome of Ythdf2-vKO and control mice were generated by using Illumina Hiseq Xten, and were further analyzed according with our previoursly reported database.