高通量测序技术发展使得大量长链编码RNA（lncRNA）被鉴定出来，同时研究表明睾丸为lncRNA特异高表达的两组织之一，指向lncRNA极可能参与精子发生过程的调控。为揭示非编码RNA介导的信息传递方式和分子机制，本项目以模式生物果蝇为代表，系统的鉴定睾丸组织特异性表达的，可能参与精子发生的lncRNA基因。通过CRISPR/phiC31技术大规模建立lncRNA敲除突变品系，并结合原位功能补偿实验系统鉴定lncRNA的生物学功能。通过对lncRNA敲除果蝇突变株进行缺陷表型分析，鉴定出在减数分裂后精子细胞核压缩、形态发生以及精子延伸中具有重要功能的lncRNA分子。针对若干具重要表型的功能性lncRNA基因（包括1株雄性不育突变体lncRNA：CR44456），结合生化和分子生物学手段研究lncRNA的分子调控机理。

Thousands of long noncoding RNAs (lncRNAs; > 200 nucleotides) have been identifiedby genome-wide transcriptome analyses. Recent studies show that lncRNAs are often predominantly transcribed in testis in both vertebrates and Drosophila, suggesting that these RNAs play similar functional roles in spermatogenesis across a broad range of animal taxa. However, it remains unclear whether lncRNAs are truly.involved in male reproduction in vivo. Thus we attempted to survey the functional roles of lncRNAs in spermatogenesis using our optimized CRISPR system. In this study, first we plan to identify hundreds of testis-specific Drosophila lncRNAs through bioinformatics prediction, published gene expression data analysis and RNA-seq. To comprehensively analyze lncRNA functions in vivo, we plan to inactivate these lncRNA genes by genomic deletion using an optimized three-component CRISPR/Cas9 system. Through quantitative fertility assay, testis.morphology examination, testis squash and DAPI staining et al., we would check phenotypes of lncRNA knockout mutants and select lncRNAs with critical function in spermatogenesis. Then we would explore the mechanism of several lncRNAs with severe phenotypes by which it may act using various methods such as RNA-seq, ChIRP-seq and RNA pull-down. These studies will provide important genetic evidence to the in vivo function and the underlying mechanism of lncRNA in Drosophila spermatogenesis.