Panoramic transcriptome analysis and functional screening of long noncoding RNAs in mouse spermatogenesis

Abstract

Long noncoding RNAs (lncRNAs) have emerged as diverse functional regulators involved in mammalian development; however, large-scale functional investigation of lncRNAs in mammalian spermatogenesis in vivo is lacking. Here, we delineated the global lncRNA expression landscape in mouse spermatogenesis and identified 968 germ cell signature lncRNAs. By combining bioinformatics and functional screening, we identified three functional lncRNAs (<i>Gm4665</i>, <i>1700027A15Rik</i>, and <i>1700052I22Rik</i>) that directly influence spermatogenesis in vivo. Knocking down <i>Gm4665</i> hampered the development of round spermatids into elongating spermatids and disrupted key spermatogenic gene expression. Mechanistically, lncRNA <i>Gm4665</i> localized in the nucleus of round spermatids and occupied the genomic regulatory region of important spermatogenic genes including <i>Ip6k1</i> and <i>Akap3</i> These findings provide a valuable resource and framework for future functional analysis of lncRNAs in spermatogenesis and their potential roles in other biological processes.