利用深度测序技术鉴定日本血吸虫 microRNAs。
Genome-wide identification of Schistosoma japonicum microRNAs using a deep-sequencing approach.
机构信息
Shanghai-MOST Key Laboratory for Disease and Health Genomics, Chinese National Human Genome Center at Shanghai, Shanghai, China.
出版信息
PLoS One. 2009 Dec 8;4(12):e8206. doi: 10.1371/journal.pone.0008206.
BACKGROUND
Human schistosomiasis is one of the most prevalent and serious parasitic diseases worldwide. Schistosoma japonicum is one of important pathogens of this disease. MicroRNAs (miRNAs) are a large group of non-coding RNAs that play important roles in regulating gene expression and protein translation in animals. Genome-wide identification of miRNAs in a given organism is a critical step to facilitating our understanding of genome organization, genome biology, evolution, and posttranscriptional regulation.
METHODOLOGY/PRINCIPAL FINDINGS: We sequenced two small RNA libraries prepared from different stages of the life cycle of S. japonicum, immature schistosomula and mature pairing adults, through a deep DNA sequencing approach, which yielded approximately 12 million high-quality short sequence reads containing a total of approximately 2 million non-redundant tags. Based on a bioinformatics pipeline, we identified 176 new S. japonicum miRNAs, of which some exhibited a differential pattern of expression between the two stages. Although 21 S. japonicum miRNAs are orthologs of known miRNAs within the metazoans, some nucleotides at many positions of Schistosoma miRNAs, such as miR-8, let-7, miR-10, miR-31, miR-92, miR-124, and miR-125, are indeed significantly distinct from other bilaterian orthologs. In addition, both miR-71 and some miR-2 family members in tandem are found to be clustered in a reversal direction model on two genomic loci, and two pairs of novel S. japonicum miRNAs were derived from sense and antisense DNA strands at the same genomic loci.
CONCLUSIONS/SIGNIFICANCE: The collection of S. japonicum miRNAs could be used as a new platform to study the genomic structure, gene regulation and networks, evolutionary processes, development, and host-parasite interactions. Some S. japonicum miRNAs and their clusters could represent the ancestral forms of the conserved orthologues and a model for the genesis of novel miRNAs.
背景
人体血吸虫病是世界上最普遍和最严重的寄生虫病之一。日本血吸虫是该病的重要病原体之一。microRNAs(miRNAs)是一组在动物中发挥重要作用的非编码 RNA,调节基因表达和蛋白质翻译。在给定的生物体中鉴定 miRNA 的全基因组是促进我们理解基因组组织、基因组生物学、进化和转录后调控的关键步骤。
方法/主要发现:我们通过深度 DNA 测序方法,从日本血吸虫不同生命周期阶段(未成熟的尾蚴和成熟的配对成虫)的两个小 RNA 文库中测序,获得了大约 1200 万个高质量的短序列,总共包含大约 200 万个非冗余标签。基于生物信息学流程,我们鉴定了 176 个新的日本血吸虫 miRNAs,其中一些在两个阶段表现出不同的表达模式。虽然 21 个日本血吸虫 miRNAs 是后生动物中已知 miRNAs 的同源物,但 Schistosoma miRNAs 的许多位置的一些核苷酸,如 miR-8、let-7、miR-10、miR-31、miR-92、miR-124 和 miR-125,确实与其他后生动物同源物有显著不同。此外,miR-71 和一些 miR-2 家族成员串联在两个基因组位置上以反转方向模型聚类,并且两个新的日本血吸虫 miRNAs 对来自同一基因组位置的正义和反义 DNA 链。
结论/意义:日本血吸虫 miRNAs 的收集可以作为研究基因组结构、基因调控和网络、进化过程、发育和宿主-寄生虫相互作用的新平台。一些日本血吸虫 miRNAs 及其簇可能代表保守同源物的原始形式,也是新 miRNA 起源的模型。