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北京生科院在環形RNA功能挖掘研究中取得進展

文章來源:北京生命科學研究院   發布時間:2019-03-22  【字號:     】  

    319日,國際學術期刊Cell Reports 在線發表了中國科學院北京生命科學研究院計算基因組學實驗室趙方慶團隊題為Expanded expression landscape and prioritization of circular RNAs in mammals 的研究成果。該研究從人、猴和小鼠三個物種的44個正常組織中鑒定出大量環形RNA,其中超過70%的環形轉錄本實現了全長重建。利用這些數據,研究人員對環形RNA的多樣性、保守性、剪接模式以及與線性RNA的成環比差異進行了多方位深入分析。研究人員還通過構建物種特異和保守的環形RNAmRNA共表達網絡,篩選具有潛在生物學功能的環形RNA。該研究為環形RNA領域的研究者提供了豐富的數據資源和高效篩選功能環形RNA的方法,對進一步深入了解環形RNA的功能和作用機制具有重要意義。

  作為一類新型的內源性非編碼RNA分子,環形RNA已經成為RNA研究中的熱點領域。雖然目前已有海量的環形RNA被識別出來,但迄今只有極少數的環形RNA分子的功能得以被揭示,其所涉及的功能主要集中在miRNA海綿、親本基因的順式調控、競爭性結合RBP和翻譯短肽等。目前,環形RNA研究領域面臨的一個關鍵問題是:如何從海量的環形RNA中高效地篩選出具有潛在生物學功能的環形RNA分子。以往的研究方法大多是利用差異表達分析,篩選出少量的候選分子,然后進行后續功能驗證。此類方法往往通量低、耗時長、效果差。物種間保守性分析也是篩選候選功能環形RNA的有效策略,但是目前的環形RNA識別算法還不能有效地識別環形RNA的全長序列,因此,其保守性分析只能局限在環形RNA的反向剪接位點附近,而無法了解其內部的保守性和剪接模式。 

  針對上述問題,趙方慶團隊采用多重文庫策略對哺乳類重要模式物種(人、猴和鼠)44個組織進行了高通量轉錄組測序,識別出大量全新的環形RNA分子(平均每個物種94461個),利用他們自主開發的CIRI-full工具對識別出的環形RNA進行全長重構,結果顯示大約有72.6%的環形RNA的全長序列得到了重建。這些結果將為人們研究環形RNA的多樣性、保守性、剪接模式以及與線性RNA的比較分析提供重要的數據基礎。利用重建的環形RNA全長序列,該團隊首次發現了一類在多個物種中高度保守的環形RNAOverlapped orthologous circRNA, OO type circRNA)。分析表明,此類OO-型環形RNA比其他類型的環形RNA擁有更高的表達量、更保守的表達模式、更高的成環比、更保守的剪接模式和功能富集。這些特征提示OO-型環形RNA可能擁有更加重要的生物學功能,更適合作為后續功能挖掘的候選。

  利用環形RNA在多個組織中的表達,該團隊首先對每個物種構建了物種特異的環形RNAmRNA的共表達網絡,隨后合并三個物種的共表達網絡進一步構建了物種間保守的共表達網絡,并對其中關鍵調控基因進行敲低實驗,以驗證該網絡的可靠性。在這些網絡的基礎上,研究人員還利用guilt-by-association原理對大部分的環形RNA進行了功能注釋。這些共表達網絡和環形RNA的保守性分析結果都為后續環形RNA的功能篩選提供了可靠的數據和方法。研究人員還將該研究中產生的數據和開發的分析方法應用至肝癌環形RNA的研究中,成功篩選出了與肝癌發生相關的環形RNA,并進行了功能驗證。最后,為了方便環形RNA研究領域的工作者方便獲取和使用該研究中的數據,研究人員建立了相關的數據及信息庫:circAtlas (http://circatlas.biols.ac.cn.)。研究人員可以快速獲取不同物種不同組織的環形RNA全長序列,確認其保守性及剪接模式,并結合網絡分析獲得其功能注釋信息。

  該工作主要由趙方慶課題組的助理研究員冀培豐和研究生吳婉瑩、陳帥共同完成,并獲得國家自然科學基金委、科技部重點研發計劃及中科院的經費支持。趙方慶團隊在前期的工作中建立了環形RNA識別、轉錄本組裝、可變剪接檢測及定量等方法,相關工作發表在Genome Biology (2015)Nature Communications (2016)Briefings in Bioinformatics (2017)Trends in Genetics (2018)Genome Medicine (2019)Cell Reports (2019)。這些研究豐富了人們對環形RNA的組成及結構的認識,為深入了解這一嶄新類型的非編碼RNA分子提供了重要方法學基礎。

  論文鏈接 

 

哺乳動物環形RNA的識別和功能挖掘 




(責任編輯:葉瑞優)

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