7月初，四川農(nóng)業(yè)大學(xué)動物遺傳育種研究所首次全面系統(tǒng)地闡述了豬從出生前到出生后整個發(fā)育階段的microRNA組（MicroRNAome）圖譜。此項研究由四川農(nóng)業(yè)大學(xué)李學(xué)偉教授（通訊作者）和李明洲博士（第一作者）領(lǐng)導(dǎo)的課題組與美國休斯頓大學(xué),，LC Sciences等機構(gòu)的科研人員合作完成,。研究成果發(fā)表于國際知名學(xué)術(shù)期刊PLoS ONE上,。
早在九千年前,，豬就已被人類馴養(yǎng)，作為重要的食物蛋白質(zhì)來源,，由于其個體大小,，解剖學(xué)，生理學(xué),，新陳代謝,，病理學(xué)以及藥理學(xué)上與人十分相近,，現(xiàn)已成為一種重要的研究人類疾病的模式動物，將來有可能成為人類異種移植的重要器官來源,。microRNA（miRNA）作為一類長度約為22 nt的非編碼小RNA,，已被大量實驗證實是一種重要的基因表達調(diào)控因子。miRNA通過抑制mRNA翻譯或是引起mRNA降解來實現(xiàn)其對基因表達的調(diào)控,。然而,，相比于人microRNAome，豬的miRNA研究嚴(yán)重滯后,。在Sanger miRBase 15.0版中,，人miRNA序列已超過900條，而豬miRNA序列僅有175條,。
為了廣泛深入地發(fā)掘豬microRNAome圖譜,，該研究收集了從胚胎到成年共計10個發(fā)育階段的樣品，覆蓋了豬發(fā)育的全部代表性時期,，采用Solexa深度測序技術(shù)分別對10份樣品進行miRNA序列的深度測定，總共獲取了超過93.6M的序列讀數(shù),。課題組使用分析軟件ACGT101-miR對測序數(shù)據(jù)進行了深度發(fā)掘,。通過與哺乳動物成熟體miRNA序列，前體發(fā)夾序列（pre-miRNA）,，首次發(fā)布的豬基因組序列（Sscrofa9,，April 2009），以及EST序列的精細比對分析,，課題組對豬microRNAome數(shù)據(jù)庫進行了大幅更新,，pre-miRNA序列增至867條，編碼的成熟體miRNA序列增至1004條,，其中特異miRNA序列達到777條,。研究人員選擇了測序得到的30條miRNA，采用實時熒光定量PCR（qPCR）對豬的47個不同類型的組織樣品進行檢測,，定量結(jié)果與測序結(jié)果一致,。科研人員對測序得到的豬microRNAome進行了詳盡的生物信息學(xué)分析,，提供了有關(guān)miRNA的末端序列變異,，miRNA前體結(jié)構(gòu)，染色體定位,，特定發(fā)育階段的miRNA表達,，以及miRNA保守性等詳細信息。
此項研究受到了國家轉(zhuǎn)基因生物新品種培育科技重大專項和國家自然科學(xué)基金的資助,，是迄今對豬microRNAome圖譜進行的最全面最詳實的探索（包括miRNA和其異構(gòu)體（isomiR））,，歸納整理了大量豬miRNA的特征,。此項研究成果的發(fā)表將大大促進豬生物學(xué)的發(fā)展，也必將促進以豬作為模式動物進行人類生物學(xué)和生物醫(yī)學(xué)的研究,。(生物谷Bioon.com)
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生物谷推薦原文出處：
PLoS ONE DOI:10.1371/journal.pone.0011541
MicroRNAome of Porcine Pre- and Postnatal Development
Mingzhou Li1#, Youlin Xia2#, Yiren Gu1, Kai Zhang1, Qiulei Lang3,4, Lei Chen5, Jiuqiang Guan1, Zonggang Luo1, Haosi Chen2, Yang Li1, Qinghai Li1, Xiang Li1, An-an Jiang1, Surong Shuai1, Jinyong Wang5, Qi Zhu4, Xiaochuan Zhou4, Xiaolian Gao2,6*, Xuewei Li1*
1 Institute of Animal Genetics and Breeding, College of Animal Science 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 and Technology of China, Hefei, Anhui, China
The domestic pig is of enormous agricultural significance and valuable models for many human diseases. Information concerning the pig microRNAome (miRNAome) has been long overdue and elucidation of this information will permit an atlas of microRNA (miRNA) regulation functions and networks to be constructed. Here we performed a comprehensive search for porcine miRNAs on ten small RNA sequencing libraries prepared from a mixture of tissues obtained during the entire pig lifetime, from the fetal period through adulthood. The sequencing results were analyzed using mammalian miRNAs, the precursor hairpins (pre-miRNAs) and the first release of the high-coverage porcine genome assembly (Sscrofa9, April 2009) and the available expressed sequence tag (EST) sequences. Our results extend the repertoire of pig miRNAome to 867 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 samples and found agreement between the sequencing and q-PCR data. This broad survey provides detailed information about multiple variants of mature sequences, precursors, chromosomal organization, development-specific expression, and conservation patterns. Our data mining produced a broad view of the pig miRNAome, consisting of miRNAs and isomiRs and a wealth of information of pig miRNA characteristics. These results are prelude to the advancement in pig biology as well the use of pigs as model organism for human biological and biomedical studies.
