三分排列3

環狀RNA芯片

  • 簡介
  • 芯片特點
  • 實驗流程
  • 客戶案例
  • SCI成果
  •        環狀RNA(circRNA)是一類由特殊剪接機制形成的、具有閉合環狀結構、大量存在于真核轉錄組中的非編碼RNA;也是目前生命科學和醫學領域的研究熱點分子。circRNA分子的組織特異性、疾病特異性、時序特異性及高穩定性等特征,使得circRNA作為臨床疾病的biomarker具有明顯的優勢。近來研究顯示,環狀RNA在不同物種中起到miRNA海綿的作用,稱之為競爭性內源RNA(ceRNA),即能競爭性結合miRNA。而與疾病關聯性miRNA的相互作用說明環狀RNA對疾病的調控起著非常重要的作用。此外,一些內含子類型的環狀RNA(ciRNA)會促進宿主基因的轉錄。
           為了能夠更好地研究circRNA,美國Arraystar公司在全球首款circRNA芯片基礎上迅即升級版本為V2.0。其circRNA來源融合了環狀RNA研究的最新頂尖文獻,所有cicrRNA都經過了嚴謹的實驗驗證,以便于對不同生理及病理條件下的circRNA進行系統的研究。同時我們對所有差異表達的circRNA用高匹配值的miRNA靶標位點進行了標注,這將有利于對circRNA作為天然miRNA海綿功能的進行研究。

    Aksomics(康成生物)是Arraystar公司中國地區唯一代理商,為您提供一站式circRNA芯片技術服務,您只需要提供保存完好的組織或細胞標本,Aksomics的技術服務人員就可為您完成全部實驗操作,并提供完整的實驗報告。

    Arraystar公司circRNA芯片產品列表


    服務 芯片 規格 描述
    Human circular RNA Microarray Service Human circular RNA Array V2.0 8x15K 13,617 human circular RNAs
    Mouse circular RNA Microarray Service Mouse circular RNA Array V2.0 8×15K 14,236 Mouse circular RNAs
    Rat circular RNA Microarray Service Rat circular RNA Array V2.0 8×15K 14,145 Rat circular RNAs
  • ● 全面、系統的circRNA收集:circRNA研究領域頂尖期刊和權威公共數據庫,運用強大的信息篩選系統,挑選出最可靠的circRNA。

    ● 高效、精準的circRNA檢測:剪接位點特異性探針與Rnase R預處理雙重保障,即便相應線性RNA存在也能特異性檢測circRNA。

    ● 嚴謹、可靠的Spike-in對照:在RNA樣品中加入一組由ERCC (External RNA Controls Consortium)開發的外源性RNA,可對RNA擴增,標記和芯片雜交過程中產生的實驗偏差進行校正,增加樣本間結果比較的可靠性。

    ● 詳盡、豐富的circRNA注釋:Arraystar miRNA靶基因預測軟件對circRNA上的保守miRNA的結合位點進行了預測,并對其中匹配分值較高的位點進行了注釋(下圖)。此外,我們還提供了與circRNA對應的線性mRNA信息。根據信息,客戶可以很方便地從不同角度研究circRNA的作用機制和生物功能。


  • 1. 樣品RNA抽提
           實驗對象為組織樣品,取適量(50-100mg)新鮮組織樣品或正確保存的組織樣品,加1ml的RNA抽提試劑TRIzol(Invitrogen),勻漿后抽提RNA。
           實驗對象為細胞樣品,每份樣品取1×106~1×107細胞,完全吸去培養液后加1ml的RNA抽提試劑TRIzol,裂解后抽提RNA。
    2.  RNA質量檢測
           使用Nanodrop測定RNA在分光光度計260nm、280nm和230nm的吸收值,以計算濃度并評估純度。
           用甲醛電泳試劑進行變性瓊脂糖凝膠電泳,檢測RNA純度及完整性。
           提供RNA QC報告。
    3. RNase R處理
    4. cDNA/aRNA樣品合成和標記
    5. 標記效率質量檢測
           使用Nanodrop檢測熒光標記效率,標記效率合格以保證后續芯片實驗結果的可靠性。
    6. 芯片雜交
           在標準條件下將標記好的探針和高密度基因組芯片進行雜交。
    7. 圖像采集和數據分析
           使用GenePix 4000B芯片掃描儀掃描芯片的熒光強度,并將實驗結果轉換成數字型數據保存,使用配套軟件對原始數據進行分析運算。
    8. 提供實驗報告:
           ? Scanning Image: Cy3熒光掃描圖像;
           ? Scatter Plot:散點圖,X軸為Control信號值,Y軸為Experiment信號值,表示信號數據總體分布趨勢;
           ? Raw Data:探針的掃描熒光信號強度原始數據;
           ? Normalized Data:經過統計學方法標準化后數值;
           ? P–value: T-test統計分析顯著差異表達的基因,p-value越小,該circRNA在兩組樣本間差異表達越顯著;        

           ? Significant Up & Down Regulated CircRNA List:顯著差異表達的circRNA列表包含Fold Change, p-value。


  • circRNA在免疫衰老中的相關研究(Comprehensive circular RNA profiling reveals that circular RNA100783 is involved in chronic CD28-associated CD8(+)T cell ageing. Immunity & Ageing, 2015)

           生物機體免疫系統隨著年齡的增長而不斷惡化,這個過程被稱為免疫衰老(immunosenescence);免疫衰老與系統炎癥反應、慢性炎癥疾病及癌癥發生等密切相關,但目前對引發免疫衰老的機制了解的甚少。該研究通過Arraystar circRNA芯片篩選發現:在健康成人與免疫力低下的老年人的外周血淋巴細胞中,circRNA的表達譜具有明顯的差異。在差異表達的circRNA中,研究者通過qPCR驗證發現hsa_circRNA_100783可以明顯區分兩種不同類型的樣本,可以作為免疫衰老的biomarker。研究者接著對circRNA可能結合的miRNA進行了靶基因預測,推測circRNA很可能通過調控磷酸蛋白信號轉導途徑來影響免疫細胞衰老的過程。

  • → A Circular RNA Protects Dormant Hematopoietic Stem Cells from DNA Sensor cGAS-Mediated Exhaustion. Immunity. 2018

    → Circular RNA MTO1 acts as the sponge of miR-9 to suppress hepatocellular carcinoma progression.  Hepatology (Baltimore, Md.). 2017

    → circRNA Mediates Silica-Induced Macrophage Activation Via HECTD1/ZC3H12A-Dependent Ubiquitination. Theranostics. 2017

    → A Circular RNA Binds To and Activates AKT Phosphorylation and Nuclear Localization Reducing Apoptosis and Enhancing Cardiac Repair.Theranostics. 2017

    → The circular RNA circPRKCI promotes tumor growth in lung adenocarcinoma. Cancer Research. 2018

    → circRNA_100290 plays a role in oral cancer by functioning as a sponge of the miR-29 family. Oncogene. 2017

    → Silencing circular RNA hsa_circ_0000977 suppresses pancreatic ductal adenocarcinoma progression by stimulating miR-874-3p and inhibiting PLK1 expression. Cancer Letters. 2018

    → Circular RNA_LARP4 inhibits cell proliferation and invasion of gastric cancer by sponging miR-424-5p and regulating LATS1 expression. Molecular Cancer. 2017

    → Circular RNA expression is suppressed by androgen receptor (AR)-regulated adenosine deaminase that acts on RNA (ADAR1) in human hepatocellular carcinoma. Cell Death & Disease. 2017

    → Comprehensive Circular RNA Profiling Reveals That hsa_circ_0005075, a New Circular RNA Biomarker, Is Involved in Hepatocellular Crcinoma Development. Medicine. 2016

    → CircRNA_000203 enhances the expression of fibrosis-associated genes by derepressing targets of miR-26b-5p, Col1a2 and CTGF, in cardiac fibroblasts. Scientific Reports. 2017

    → Circular RNA Related to the Chondrocyte ECM Regulates MMP13 Expression by Functioning as a MiR-136 'Sponge' in Human Cartilage Degradation. Scientific Reports. 2016

    → Circular RNA GRB10 as a competitive endogenous RNA regulating nucleus pulposus cells death in degenerative intervertebral disk. Cell Death & Disease. 2018

    → circGFRA1 and GFRA1 act as ceRNAs in triple negative breast cancer by regulating miR-34a. Journal of Experimental & Clinical Cancer Research. 2017

    → Circ100284, via miR-217 regulation of EZH2, is involved in the arsenite-accelerated cell cycle of human keratinocytes in carcinogenesis. Biochimica et Biophysica Acta (BBA)-Molecular Basis of Disease. 2017

    → CircRNA Expression Profile in Early-Stage Lung Adenocarcinoma Patients. Cellular Physiology and Biochemistry. 2017

    → Altered Circular RNA Expression in Patients with Repeated Implantation Failure. Cellular Physiology and Biochemistry. 2017

    → Microarray Expression Profile and Functional Analysis of Circular RNAs in Osteosarcoma. Cellular Physiology and Biochemistry. 2017

    → Microarray Expression Profile of Circular RNAs in Peripheral Blood Mononuclear Cells from Rheumatoid Arthritis Patients.Cellular Physiology and Biochemistry. 2017

    → Circular RNA circ-4099 is induced by TNF-α and regulates ECM synthesis by blocking miR-616-5p inhibition of Sox9 in intervertebral disc degeneration. Experimental & molecular medicine. 2018