title: Critical roles of long noncoding RNAs in Drosophila spermatogenesis.
abstract: Long noncoding RNAs (lncRNAs), a recently discovered class of cellular RNAs, play important roles in the regulation of many cellular developmental processes. Although lncRNAs have been systematically identified in various systems, most of them have not been functionally characterized in vivo in animal models. In this study, we identified 128 testis-specific Drosophila lncRNAs and knocked out 105 of them using an optimized three-component CRISPR/Cas9 system. Among the lncRNA knockouts, 33 (31%) exhibited a partial or complete loss of male fertility, accompanied by visual developmental defects in late spermatogenesis. In addition, six knockouts were fully or partially rescued by transgenes in a trans configuration, indicating that those lncRNAs primarily work in trans Furthermore, gene expression profiles for five lncRNA mutants revealed that testis-specific lncRNAs regulate global gene expression, orchestrating late male germ cell differentiation. Compared with coding genes, the testis-specific lncRNAs evolved much faster. Moreover, lncRNAs of greater functional importance exhibited higher sequence conservation, suggesting that they are under constant evolutionary selection. Collectively, our results reveal critical functions of rapidly evolving testis-specific lncRNAs in late Drosophila spermatogenesis.
authors: MOE Key Laboratory of Bioinformatics, Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing 100084, China;
authors: Kejia
authors: K
authors: Wen
authors: MOE Key Laboratory of Bioinformatics, Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing 100084, China; College of Animal Science, Tarim University, Xinjiang 843300, China;
authors: Lijuan
authors: L
authors: Yang
authors: MOE Key Laboratory of Bioinformatics, Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing 100084, China;
authors: Tuanlin
authors: T
authors: Xiong
authors: MOE Key Laboratory of Bioinformatics, Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing 100084, China;
authors: Chao
authors: C
authors: Di
authors: MOE Key Laboratory of Bioinformatics, Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing 100084, China;
authors: Danhui
authors: D
authors: Ma
authors: MOE Key Laboratory of Bioinformatics, Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing 100084, China;
authors: Menghua
authors: M
authors: Wu
authors: MOE Key Laboratory of Bioinformatics, Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing 100084, China;
authors: Zhaoyu
authors: Z
authors: Xue
authors: MOE Key Laboratory of Bioinformatics, Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing 100084, China;
authors: Xuedi
authors: X
authors: Zhang
authors: MOE Key Laboratory of Bioinformatics, Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing 100084, China;
authors: Li
authors: L
authors: Long
authors: MOE Key Laboratory of Bioinformatics, Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing 100084, China;
authors: Weimin
authors: W
authors: Zhang
authors: MOE Key Laboratory of Bioinformatics, Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing 100084, China;
authors: Jiaying
authors: J
authors: Zhang
authors: Institute of Cancer Stem Cell, Dalian Medical University, Dalian 116044, China;
authors: Xiaolin
authors: X
authors: Bi
authors: MOE Key Laboratory of Bioinformatics, Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing 100084, China; Center for Synthetic and Systems Biology, Tsinghua University, Beijing 100084, China;
authors: Junbiao
authors: J
authors: Dai
authors: MOE Key Laboratory of Bioinformatics, Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing 100084, China; Center for Synthetic and Systems Biology, Tsinghua University, Beijing 100084, China; Beijing Advanced Innovation Center for Structural Biology, Tsinghua University, Beijing 100084, China.
authors: Qiangfeng
authors: Q
authors: Zhang
authors: MOE Key Laboratory of Bioinformatics, Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing 100084, China; Center for Synthetic and Systems Biology, Tsinghua University, Beijing 100084, China;
authors: Zhi John
authors: ZJ
authors: Lu
authors: MOE Key Laboratory of Bioinformatics, Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing 100084, China; Center for Synthetic and Systems Biology, Tsinghua University, Beijing 100084, China;
authors: Guanjun
authors: G
authors: Gao
mesh_terms: D000818:Animals; D064113:CRISPR-Cas Systems; D017124:Conserved Sequence / Q000235:genetics*; D004330:Drosophila / Q000235:genetics / Q000254:growth & development; D018507:Gene Expression Regulation, Developmental; D005854:Germ Cells / Q000254:growth & development; D007248:Infertility, Male / Q000235:genetics / Q000473:pathology; D008297:Male; D062085:RNA, Long Noncoding / Q000235:genetics*; D013091:Spermatogenesis / Q000235:genetics*; D013737:Testis / Q000254:growth & development*
chemical_list: D062085:RNA, Long Noncoding