Identification and characterization of novel microRNAs from Schistosoma japonicum.
Identification and characterization of novel microRNAs from Schistosoma japonicum.
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DOI：
10.1371/journal.pone.0004034
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发表时间：
2008
期刊：
影响因子：
3.7
通讯作者：
Pan W
中科院分区：
文献类型：
--
作者：
Xue X;Sun J;Zhang Q;Wang Z;Huang Y;Pan W
日本血吸虫病仍然是中国的主要公共卫生问题。其病原体日本血吸虫具有复杂的生命周期和在不同生命周期阶段表达的独特基因库。探索复杂的基因调控将为新的药物靶点和候选疫苗带来最好的前景。microRNA（miRNAs）是一类高度保守的非编码RNA，通过序列特异性抑制基因表达来调控多种生物学过程。尽管从植物到哺乳动物已经鉴定出大量的miRNAs，但是否存在miRNAs的研究还没有实验证据。本研究通过对日本血吸虫成虫小RNA（18-26 nt）cDNA文库的克隆和测序，从日本血吸虫中鉴定出了新的miRNAs。从227个克隆的RNA序列中鉴定出5个新的miRNAs，并通过北方杂交进行验证。miRNA与相应家族成员的比对表明，其中四个属于后生动物miRNA家族：let-7，miR-71，bantam和miR-125。第五个潜在的新（非保守）miRNA似乎属于血吸虫属中以前未描述的家族。新发现的miRNAs分别命名为sja-let-7、sja-miR-71、sja-bantam、sja-miR-125和sja-miR-new 1。采用本实验室改进的茎环逆转录聚合酶链反应（RT-PCR）方法，分析了Sja-let-7、Sja-miR-71和Sja-bantam在虫卵、毛蚴、孢子囊、尾蚴、童虫和成虫等6个阶段的表达。这些miRNA的表达模式具有高度的阶段特异性。特别地，sja-miR-71和sja-bantam表达在尾蚴阶段达到其峰值，然后在尾蚴渗透到哺乳动物宿主中后，在幼体阶段快速下降至最低点。首次在S. japonicum，包括一个新的双体家族成员。新的miRNAs在不同的生命阶段的不同表达模式。推测它们可能在血吸虫的生长发育中起重要作用。
Schistosomiasis japonica remains a major public health problem in China. Its pathogen, Schistosoma japonicum has a complex life cycle and a unique repertoire of genes expressed at different life cycle stages. Exploring schistosome gene regulation will yield the best prospects for new drug targets and vaccine candidates. MicroRNAs (miRNAs) are a highly conserved class of noncoding RNA that control many biological processes by sequence-specific inhibition of gene expression. Although a large number of miRNAs have been identified from plants to mammals, it remains no experimental proof whether schistosome exist miRNAs. We have identified novel miRNAs from Schistosoma japonicum by cloning and sequencing a small (18–26 nt) RNA cDNA library from the adult worms. Five novel miRNAs were identified from 227 cloned RNA sequences and verified by Northern blot. Alignments of the miRNAs with corresponding family members indicated that four of them belong to a metazoan miRNA family: let-7, miR-71, bantam and miR-125. The fifth potentially new (non conserved) miRNA appears to belong to a previously undescribed family in the genus Schistosome. The novel miRNAs were designated as sja-let-7, sja-miR-71, sja-bantam, sja-miR-125 and sja-miR-new1, respectively. Expression of sja-let-7, sja-miR-71 and sja-bantam were analyzed in six stages of the life cycle, i.e. egg, miracidium, sporocyst, cercaria, schistosomulum, and adult worm, by a modified stem-loop reverse transcribed polymerase chain reaction (RT-PCR) method developed in our laboratory. The expression patterns of these miRNAs were highly stage-specific. In particular, sja-miR-71 and sja-bantam expression reach their peaks in the cercaria stage and then drop quickly to the nadirs in the schistosomulum stage, following penetration of cercaria into a mammalian host. Authentic miRNAs were identified for the first time in S. japonicum, including a new schistosome family member. The different expression patterns of the novel miRNAs over the life stages of S. japonicum suggest that they may mediate important roles in Schistosome growth and development.
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作者：
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