Center for RNA Biology: From Genome to Therapeutics, University of Rochester Medical Center, Rochester, NY, USA.
Department of Biology, University of Rochester, Rochester, NY, USA.
Nat Commun. 2021 Mar 1;12(1):1361. doi: 10.1038/s41467-021-21524-6.
Sperm contributes diverse RNAs to the zygote. While sperm small RNAs have been shown to impact offspring phenotypes, our knowledge of the sperm transcriptome, especially the composition of long RNAs, has been limited by the lack of sensitive, high-throughput experimental techniques that can distinguish intact RNAs from fragmented RNAs, known to abound in sperm. Here, we integrate single-molecule long-read sequencing with short-read sequencing to detect sperm intact RNAs (spiRNAs). We identify 3440 spiRNA species in mice and 4100 in humans. The spiRNA profile consists of both mRNAs and long non-coding RNAs, is evolutionarily conserved between mice and humans, and displays an enrichment in mRNAs encoding for ribosome. In sum, we characterize the landscape of intact long RNAs in sperm, paving the way for future studies on their biogenesis and functions. Our experimental and bioinformatics approaches can be applied to other tissues and organisms to detect intact transcripts.
精子向受精卵贡献了多种 RNA。虽然已经证实精子中的小 RNA 会影响后代的表型，但由于缺乏敏感、高通量的实验技术，我们对精子转录组的了解，特别是长 RNA 的组成，一直受到限制，这些技术可以区分完整的 RNA 和已知在精子中大量存在的片段化 RNA。在这里，我们将单分子长读测序与短读测序相结合，以检测精子完整 RNA（spiRNA）。我们在小鼠中鉴定出 3440 种 spiRNA 物种，在人类中鉴定出 4100 种 spiRNA 物种。spiRNA 图谱由 mRNA 和长非编码 RNA 组成，在小鼠和人类之间具有进化保守性，并显示出富含核糖体编码 mRNA 的特征。总之，我们描述了精子中完整长 RNA 的图谱，为进一步研究其生物发生和功能奠定了基础。我们的实验和生物信息学方法可应用于其他组织和生物体，以检测完整的转录本。
