一种新发现的、睾丸高表达的非编码RNA,RHNO1在紫外线引起的DNA双链断裂修复过程中的作用

RHNO1: A novel noncoding RNA highly expressed in the testis and involved in DNA double-strand break repair

目的:发现一个转录来自RHNO1基因的长链非编码RNA(lncRNA)并分析其在脱氧核糖核酸双链断裂(DSB)修复中的功能。方法:以过表达质粒为基础,运用免疫印迹、Real-time PCR和LC/MSMS共同验证RHNO1基因的转录、翻译情况。通过Real-time PCR分析了RHNO1在小鼠各组织中的转录水平的表达情况。通过原位杂交法分析RHNO1在生精周期中的表达情况。以UV照射引起的生殖细胞系DSB为模型,进一步验证RHNO1在DSB修复中的作用。结果:全长RHNO1基因可以表达lncRNA。RHNO1基因在小鼠睾丸组织中表达最高并主要定位在精母细胞和圆形精子中。RHNO1在生殖细胞系中具有DSB修复作用。结论:发现了新的长链非编码RNA,RHNO1,其在小鼠睾丸组织中高表达,并在生殖细胞系中具有DNA损伤修复作用。

Objective: To investigate the role of a long noncoding RNA(lncRNA) transcribed from the RHNO1 gene we newly identified in DNA double-strand break(DSB) repair. Methods: The transcription and translation of the RHNO1 gene were validated by Western blot, real-time PCR and liquid chromatography-tandem mass spectrometry(LC-MS/MS) based on the overexpressed RHNO1 plasmid. The transcription level of RHNO1 in the mouse tissue was detected by real-time PCR and its expression in the spermatogenic cycle determined by in situ hybridization. The role of RHNO1 in the DNA DSB repair was further verified using the DSB model established by exposing the germ cells to ultraviolent radiation. Results: The full-length RHNO1 gene could be transcribed as a novel lncRNA in vitro, highly expressed in the mouse testis tissue, and mainly located in spermatocytes and round spermatids. RHNO1 was involved in DNA DSB repair in the spermatogenic cells. Conclusion: We identified a novel lncRNA, RHNO1, which is highly expressed in the mouse testis and participates in DNA damage repair in the germ cell line.