PLoS ONE：猪完整发育阶段microRNAome图谱发布-生物研究-生物谷

7月初，四川农业大学动物遗传育种研究所首次全面系统地阐述了猪从出生前到出生后整个发育阶段的microRNA组（MicroRNAome）图谱。此项研究由四川农业大学李学伟教授（通讯作者）和李明洲博士（第一作者）领导的课题组与美国休斯顿大学，LCSciences等机构的科研人员合作完成。研究成果发表于国际知名学术期刊 PLoS ONE 上。

早在九千年前，猪就已被人类驯养，作为重要的食物蛋白质来源，由于其个体大小，解剖学，生理学，新陈代谢，病理学以及药理学上与人十分相近，现已成为一种重要的研究人类疾病的模式动物，将来有可能成为人类异种移植的重要器官来源。microRNA（miRNA）作为一类长度约为22nt的非编码小RNA，已被大量实验证实是一种重要的基因表达调控因子。miRNA通过抑制mRNA翻译或是引起mRNA降解来实现其对基因表达的调控。然而，相比于人microRNAome，猪的miRNA研究严重滞后。在Sanger miRBase15.0版中，人miRNA序列已超过900条，而猪miRNA序列仅有175条。

为了广泛深入地发掘猪microRNAome图谱，该研究收集了从胚胎到成年共计10个发育阶段的样品，覆盖了猪发育的全部代表性时期，采用Solexa深度测序技术分别对10份样品进行miRNA序列的深度测定，总共获取了超过93.6M的序列读数。课题组使用分析软件ACGT101-miR对测序数据进行了深度发掘。通过与哺乳动物成熟体miRNA序列，前体发夹序列（pre-miRNA），首次发布的猪基因组序列（Sscrofa9，April2009），以及EST序列的精细比对分析，课题组对猪microRNAome数据库进行了大幅更新，pre-miRNA序列增至867条，编码的成熟体miRNA序列增至1004条，其中特异miRNA序列达到777条。研究人员选择了测序得到的30条miRNA，采用实时荧光定量PCR（qPCR）对猪的47个不同类型的组织样品进行检测，定量结果与测序结果一致。科研人员对测序得到的猪microRNAome进行了详尽的生物信息学分析，提供了有关miRNA的末端序列变异，miRNA前体结构，染色体定位，特定发育阶段的miRNA表达，以及miRNA保守性等详细信息。

此项研究受到了国家转基因生物新品种培育科技重大专项和国家自然科学基金的资助，是迄今对猪microRNAome图谱进行的最全面最详实的探索（包括miRNA和其异构体（isomiR）），归纳整理了大量猪miRNA的特征。此项研究成果的发表将大大促进猪生物学的发展，也必将促进以猪作为模式动物进行人类生物学和生物医学的研究。(生物谷Bioon.com) 

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PLoS ONE DOI:10.1371/journal.pone.0011541

MicroRNAome of Porcine Pre- and Postnatal Development
MingzhouLi1#, Youlin Xia2#, Yiren Gu1, Kai Zhang1, Qiulei Lang3,4, Lei Chen5,Jiuqiang Guan1, Zonggang Luo1, Haosi Chen2, Yang Li1, Qinghai Li1, XiangLi1, An-an Jiang1, Surong Shuai1, Jinyong Wang5, Qi Zhu4, XiaochuanZhou4, Xiaolian Gao2,6*, Xuewei Li1*

1 Institute of Animal Genetics and Breeding, College of AnimalScience and Technology, Sichuan Agricultural University, Ya'an, Sichuan,China, 2 Department of Biology and Biochemistry, University of Houston,Houston, Texas, United States of America, 3 Department of Biology,Zhejiang University, Hangzhou, Zhejiang, China, 4 LC Sciences, Houston,Texas, United States of America, 5 Chongqing Academy of Animal Science,Chongqing, China, 6 School of Life Science, University of Science andTechnology of China, Hefei, Anhui, China

The domestic pig is of enormous agricultural significance andvaluable models for many human diseases. Information concerning the pigmicroRNAome (miRNAome) has been long overdue and elucidation of thisinformation will permit an atlas of microRNA (miRNA) regulationfunctions and networks to be constructed. Here we performed acomprehensive search for porcine miRNAs on ten small RNA sequencinglibraries prepared from a mixture of tissues obtained during the entirepig lifetime, from the fetal period through adulthood. The sequencingresults were analyzed using mammalian miRNAs, the precursor hairpins(pre-miRNAs) and the first release of the high-coverage porcine genomeassembly (Sscrofa9, April 2009) and the available expressed sequence tag(EST) sequences. Our results extend the repertoire of pig miRNAome to867 pre-miRNAs (623 with genomic coordinates) encoding for 1,004 miRNAs,of which 777 are unique. We preformed real-time quantitative PCR(q-PCR) experiments for selected 30 miRNAs in 47 tissue-specific samplesand found agreement between the sequencing and q-PCR data. This broadsurvey provides detailed information about multiple variants of maturesequences, precursors, chromosomal organization, development-specificexpression, and conservation patterns. Our data mining produced a broadview of the pig miRNAome, consisting of miRNAs and isomiRs and a wealthof information of pig miRNA characteristics. These results are preludeto the advancement in pig biology as well the use of pigs as modelorganism for human biological and biomedical studies.