microRNA(miRNA)属于small ncRNAs,是一类大小为21-23nt的单链短RNA分子。miRNA是基因表达的主要调节分子。miRNA以序列互补配对的方式与特异靶mRNA 的3’UTR结合,通过降解靶mRNA或YZ其蛋白翻译调控基因的表达。miRNA不但在基本的生物学过程,如发育、应激反应、代谢和基因组完整性维持等方面有基本而重要的调控作用;在疾病的发SF展全过程中,也发挥着重要的调控作用。
安捷伦(Agilent)公司凭借先进的芯片生产工艺开发出高性能miRNA芯片平台,该平台结合了独特的miRNA直接标记方法以及利用创新的SurePrint喷墨合成技术设计探针等优点,同时基于ZX版本的miRBase 21.0数据库,具有高特异性和高灵敏度的特点,能特异性地检测成熟体miRNA,并能有效区分序列高度相似的不同miRNA,可广泛应用于多种类型样品miRNA表达的检测,准确的阐明miRNA在研究中的重要作用,国内外利用该平台已经发表了大量文章。
康成生物为您提供Agilent miRNA表达谱芯片技术服务,您只需要提供保存完好的组织或细胞标本,康成生物的芯片技术服务人员就可为您完成全部实验操作,并提供完整的实验报告。同时,根据您的研究需要,康成生物还提供多平台联合分析、分子标志物筛选分析等各种深入数据挖掘服务。 |
Agilent miRNA芯片产品列表 芯片名称 | 物种 | P/N | Design ID | 规格 | 描述 |
Agilent Human miRNA Microarray, Release 21.0 | 人类 | G4872A | 070156 | 8 x 60K | 2,549 human miRNAs represented miRBase database (Release 21.0) Agilent 60-mer SurePrint technology |
Mouse miRNA Microarray, Release 21.0 | 小鼠 | G4872A | 070155 | 8 x 60K | 1,881 mouse miRNAs represented miRBase database (Release 21.0) Agilent 60-mer SurePrint technology |
Rat miRNA Microarray, Release 21.0 | 大鼠 | G4471A | 070154 | 8 x 15K | 758 rat miRNAs represented miRBase database (Release 21.0) Agilent 60-mer SurePrint technology |
Agilent miRNA芯片特点1.创新的标记系统和探针设计:GX的直接标记方法,独特的SurePrint原位喷墨合成技术合成60-mer寡核苷酸探针,可以方便的区分成熟miRNA和miRNA前体,同时可以为序列高度相似的不同miRNA检测提供ZJ的灵敏度和特异性。
Components of the Agilent miRNA microarray probe design
2.数据来源于Sanger miRBase 21.0ZX版本,芯片数据更新最迅速,为客户提供最前沿的miRNA研究信息。
3.灵敏度高,只需100ng的Total RNA即可用来进行标记实验。
4.动态范围广,检测丰度跨4个数量级,有利于检测到更宽丰度范围的miRNA。
5.样品适用范围广。
6.灵活方便的定制服务。
康成生物miRNA芯片技术服务实验流程1.样品RNA抽提
2.RNA质量检测
3.制备荧光标记探针,miRNA 3’端进行Cy荧光标记,采用SurePrint原位喷墨合成技术合成60-mer用于与芯片杂交的荧光探针。
4.芯片杂交,取一定量的质检达标样本,与miRNA芯片进行杂交。
5.图像采集和数据分析。
6.提供实验报告—包括详细的实验方法和芯片实验数据及图表。
康成生物miRNA芯片技术服务基本数据分析结果展示1. miRNA差异表达数据 通过芯片中位值对miRNA芯片原始信号值进行标准化。通过标准化信号值计算出每个miRNA在不同样品间的表达变化(fold change),并通过t-test计算样品间miRNA表达量显著性p值。针对多重比较,p值被校正为FDR。根据倍数变化,p值或者FDR等参数筛选差异表达的miRNA。
适用范围:两个或两组样品间的比较,建议每组样品数目大于或等于3。
图释:样品间差异表达的miRNA数据示例。2. 差异表达miRNA的聚类图 层次聚类是一种最常见的用于分析表达数据的聚类方法。它可以根据样品中基因的表达水平将样品自动的分组,可以让客户从整体上评估样品间的基因表达差异,以及样品间的关系。左侧的树状图可以反映样品间基因表达模式的关系。
适用范围:两组或多组样品的miRNA表达谱分析
图释:不同样品组之间差异表达miRNA的聚类示例。
3. 差异表达miRNA的火山图 火山图可以对不同样品组之间差异表达的miRNA进行图形化的展示,直观的展示miRNA在样品组间的倍数变化与相应的统计学显著性之间的关系。横轴代表差异miRNA表达倍数的变化(log2转化),纵轴代表相应的p值(-log10转化)。
适用范围:两组样品间的差异miRNA分析,每组样品数目必须大于等于3。
图释:火山图示例。
康成生物miRNA芯片技术服务高级数据分析结果展示1.差异表达miRNA的靶基因预测及miRNA-靶基因网路图构建 康成生物通过三大数据库(miranda, mirbase, targetscan)预测miRNA靶点信息,将三种数据库的结果交集作为ZZmiRNA的靶基因结果,有效的降低了靶基因预测的假阳性率。Venn图展示了对于特定miRNA列表利用上述数据库的多个预测结果的整合分析。同时构建了miRNA-靶基因的网络图,直观的展示miRNA及其靶基因的调控关系。
适用范围:两组或多组数据比较获得的差异miRNA,一般情况下用于网络分析的miRNA数目应小于10个。
图释:左图:康成生物miRNA靶基因预测结果的Venn图示例;右图:康成生物miRNA—靶基因网络图示例。2.靶基因的功能分析:Go & Pathway分析 为了获得对miRNA生物学功能的理解,鉴定其全部靶mRNA(靶基因)是必不可少的。康成使用多种数据库来预测miRNA靶基因。之后对这些基因的功能进行分析(GO分析和KEGG pathway分析)。
图释:A. GO分析结果展示:预测的差异表达miRNA靶基因的前10个富集生物学过程;B. Pathway分析结果展示:预测的miRNA靶基因(黄色标注)富集到的细胞周期相关通路。康成生物miRNA芯片与其它产品联合数据分析结果展示1.miRNA芯片与lncRNA/circRNA芯片结果联合分析(ceRNA Analysis) 康成生物通过MuTaMe(mutually targeted MRE enrichment )分析寻找我们感兴趣的lncRNA/circRNA的ceRNAs,进而构建miRNA、lncRNA/circRNA和mRNA的调控网络,从而揭示ceRNA通过竞争性结合miRNA来调控目的lncRNA/circRNA的新的调控模式。
适用范围:同时具有miRNA芯片数据和lncRNA/circRNA芯片数据的样品。
图释:ceRNA分析结果展示。左侧:ceRNA网络图。右侧:针对网络图中的特定lncRNA/circRNA(lincRNA-GPR180)的lncRNA/circRNA-miRNA-mRNA相互作用详细信息。2.miRNA芯片与mRNA芯片结果联合分析 根据表达谱芯片结果和miRNA芯片结果联合分析,进一步定位差异miRNA发挥功能的途径以及相应的差异表达靶基因可能引起的生物学通路变化。
适用范围:同时具有mRNA表达谱数据和miRNA表达谱数据的样品,一般情况下用于分析的差异miRNA数目应小于10个。
图释:康成生物miRNA芯片与mRNA芯片联合分析示例:miRNA-靶基因网络图。3.miRNA与DNA甲基化芯片结果联合分析 DNA甲基化和miRNA调控是两种重要的表观遗传现象。研究表明,特定miRNA能够作为肿瘤YZmiRNA靶向DNA甲基转移酶。联合miRNA和DNA甲基化芯片结果能够研究两者之间的复杂调控关系,更好地理解癌症发生的机制。
适用范围:同时具有miRNA表达谱数据和甲基化谱数据的样品。
Agilent miRNA芯片应用案例展示筛选评估原发性骨关节炎(OA)风险和进展的潜在miRNA生物标志物(Serum microRNA array analysis identifies miR-140-3p, miR-33b-3p and miR-671-3p as potential osteoarthritis biomarkers involved in metabolic processes. Clin Epigenetics,2017) 骨关节炎(OA)是最常见的慢性退行性关节病,是导致疼痛和残疾的主要原因。了解OA发病机制涉及的分子机制对于OA个性化ZL的发现是必不可少的。然而,早期检测疾病缺乏以及开发有效的OAZL仍然是一个主要挑战。迄今为止,OA疾病诊断和严重程度判断的标准方法是X线片,似乎仅限于疾病晚期的检测,并且在监测疾病进展方面有弱点。因此,无创和敏感的血清生物标志物的鉴定将有助于OA的诊断,预后和早期ZL,
在这项研究中,我们的目标是确定OA患者的miRNA生物标志物。我们使用Agilent Human miRNA Microarray在12名原发性OA患者血清中鉴定出279个差异表达的miRNA。 ROC分析显示77个miRNA的AUC> 0.8,p <0.05。对77种miRNA进行生物信息学分析表明,它们的靶基因参与了与OA相关联的多个信号通路。选择7个排名靠前的miRNA在OA患者的血清中进行qRT-PCR验证,发现3个显著下调3倍的miRNA(hsa-miR-33b-3p, hsa-miR-671-3p和hsa-miR-140-3p),靶基因分析表明,InsR和IGFR1是3个miRNA的共同靶基因,它们主要参与OA相关的代谢过程的调控。ZZ表明我们在OA患者的血清中鉴定了三个miRNA(hsa-miR-140-3p,hsa-miR-671-3p和hsa-miR-33b-3p)可以用作评估的OA风险和进展的一个潜在的生物标志物。
技术路线结果展示图释:Agilent Human miRNA芯片筛选差异表达的miRNAs上下调统计柱形图。图释:ROC分析筛选OA相关生物标志物。图释:qPCR验证、靶基因预测及生信分析确定hsa-miR-140-3p,hsa-miR-671-3p和hsa-miR-33b-3p)用作评估的OA风险和进展的潜在的生物标志物。康成客户发表的部分SCI文章(使用康成生物miRNA芯片技术服务)1. Mutant Runx2 regulates amelogenesis and osteogenesis through a miR-185-5p-Dlx2 axis. Cell Death & Disease,2017
2. Involvement of miR-451 in resistance to paclitaxel by regulating YWHAZ in breast cancer. Cell Death and Disease,2017
3. MiRNA Expression Profile of the Myocardial Tissue of Pigs with Coronary Microembolization. Cellular Physiology and Biochemistry,2017
4. Identification of miRNA-7 by genome-wide analysis as a critical sensitizer for TRAIL-induced apoptosis in glioblastoma cells. Nucleic acids research,2017
5. Combination of AAV-TRAIL with miR-221-Zip Therapeutic Strategy Overcomes the Resistance to TRAIL Induced Apoptosis in Liver Cancer. Theranostics,2017
6. Tumor-suppressive miR-26a and miR-26b inhibit cell aggressiveness by regulating FUT4 in colorectal cancer. Cell Death & Disease,2017
7. SOX2 regulates multiple malignant processes of breast cancer development through the SOX2/miR-181a-5p, miR-30e-5p/TUSC3 axis. Molecular Cancer,2017
8. Exosomal miR-24-3p impedes T-cell function by targeting FGF11 and serves as a potential prognostic biomarker for nasopharyngeal carcinoma. J Pathol,2016
9. Exploring Transcription Factors-microRNAs Co-regulation Networks in Schizophrenia. Schizophrenia Bulletin,2016
10. Increased Variability of Genomic Transcription in Schizophrenia. Scientific Reports,2016
11. microRNA-802/Rnd3 pathway imposes on carcinogenesis and metastasis of fine particulate matter exposure. Oncotarget,2016
12. Reprogramming of Normal Fibroblasts into Cancer-Associated Fibroblasts by miRNAs-Mediated CCL2/VEGFA Signaling. PLos Genetics,2016
13. Role of microRNA-4516 involved autophagy associated with exposure to fine particulate matter. Oncotarget,2016
14. Converging Evidence Implicates the Abnormal MicroRNA System in Schizophrenia. Schizophrenia Bulletin,2015
15. Methylation-induced loss of miR-484 in microsatellite-unstable colorectal cancer promotes both viability and IL-8 production via CD137L. J Pathol,2015
16. MicroRNA-374b Suppresses Proliferation and Promotes Apoptosis in T-cell Lymphoblastic Lymphoma by Repressing AKT1 and Wnt-16. Clin Cancer Res,2015
17. Deregulated microRNAs in gastric cancer tissue-derived mesenchymal stem cells: novel biomarkers and a mechanism for gastric cancer. Brit J Cancer,2014
18. Epigenetic silencing of microRNA-199b-5p is associated with acquired chemoresistance via activation of JAG1-Notch1 signaling in ovarian cancer. Oncotarget,2014
19. miR-1207-5p and miR-1266 suppress gastric cancer growth and invasion by targeting telomerase reverse transcriptase. CELL DEATH & DISEASE,2014
20. MiR-139-5p inhibits HGTD-P and regulates neuronal apoptosis induced by hypoxia–ischemia in neonatal rats. Neurobiology of Disease,2014
21. Oncogenic miR-20a and miR-106a enhance the invasiveness of human glioma stem cells by directly targeting TIMP-2. Oncogene,2014
22. Exosomes mediate the cell-to-cell transmissi on of IFN-α-induced antiviral activity. Nature Immunology,2013
23. Identification of recurrence-related microRNAs in hepatocellular carcinoma following liver transplantation. Molecular Oncology,2013
24. MicroRNA-140-5p Suppresses Tumor Growth and Metastasis by Targeting Transforming Growth Factor β Receptor 1 and Fibroblast Growth Factor 9 in Hepatocellular Carcinoma. Hepatology,2013
25. TGF-b1 suppression of microRNA-450b-5p expression: a novel mechanism for blocking myogenic differentiation of rhabdomyosarcoma. Oncogene,2013
26. Glucocorticoids inhibit lipopolysaccharide-mediated inflammatory response by downregulating microRNA-155: a novel anti-inflammation mechanism. Free Radical Biology & Medicine,2012
27. The MicroRNA-328 Regulates Hypoxic Pulmonary Hypertension by Targeting at Insulin Growth Factor 1 Receptor and L-Type Calcium Channel- 1C. hypertensionaha,2012
28. MicroRNA-494 Is Required for the Accumulation and Functions of Tumor-Expanded Myeloid-Derived Suppressor Cells via Targeting of PTEN. The Journal of Immunology,2012
29. Deregulated miR-155 promotes Fas-mediated apoptosis in human intervertebral disc degeneration by targeting FADD and caspase-3. J Pathol,2011
30. Dysregulated expression of miR-146a contributes to agerelated dysfunction of macrophages. Anatomical Society of Great Britain and Ireland,2011
31. MicroRNA 345, a methylation-sensitive microRNA is involved in cell proliferation and invasion in human colorectal cancer. Carcinogenesis,2011
32. MicroRNA-135a contributes to the development of portal vein tumor thrombus by promoting metastasis in hepatocellular carcinoma.Shupeng . Journal of Hepatology,2011
33. Overexpression of miR-125b, a novel regulator of innate immunity, in eosinophilic chronic rhinosinusitis with nasal polyps. Am J Respir Crit Care Med,2011
34. Signature microRNA Expression Profile of Essential Hypertension and Its Novel Link to Human Cytomegalovirus Infection. Circulation,2011
35. Up-regulation of miR-21 mediates resistance to trastuzumab therapy for breast cancer. Journal of Biological Chemistry ,2011
36. MicroRNA-125b Confers the Resistance of Breast Cancer Cells to Paclitaxel through Suppression of Pro-apoptotic Bcl-2 Antagonist Killer 1 (Bak1) Expression. J Biol Chem,2010
37. MiR-218 Inhibits Invasion and Metastasis of Gastric Cancer by Targeting the Robo1 Receptor. Plos Genetics,2010
38. miR-15b and miR-16 are implicated in activation of the rat hepatic stellate cell: An essential role for apoptosis. J Hepatol,2009
39. Repression of the miR-17-92 cluster by p53 has an important function in hypoxia-induced apoptosis. EMBO J,2009
40. MicroRNA expression and regulation in mouse uterus during embryo implantation. J Biol Chem,2008