环状rna芯片

  • 简介
  • 芯片特点
  • 实验流程
  • 客户案例
  • sci成果
  •        环状rna(circrna)是一类由特殊剪接机制形成的、具有闭合环状结构、大量存在于真核转录组中的非编码rna;也是目前生命科学和医学领域的研究热点分子。circrna分子的组织特异性、疾病特异性、时序特异性及高稳定性等特征,使得circrna作为临床疾病的biomarker具有明显的优势。近来研究显示,环状rna在不同物种中起到mirna海绵的作用,称之为竞争性内源rna(cerna),即能竞争性结合mirna。而与疾病关联性mirna的相互作用说明环状rna对疾病的调控起着非常重要的作用。此外,一些内含子类型的环状rna(cirna)会促进宿主基因的转录。
           为了能够更好地研究circrna,美国arraystar公司在全球首款circrna芯片基础上迅即升级版本为v2.0。其circrna来源融合了环状rna研究的最新顶尖文献,所有cicrrna都经过了严谨的实验验证,以便于对不同生理及病理条件下的circrna进行系统的研究。同时我们对所有差异表达的circrna用高匹配值的mirna靶标位点进行了标注,这将有利于对circrna作为天然mirna海绵功能的进行研究。

    康成生物是arraystar公司中国地区唯一代理商,为您提供一站式circrna芯片技术服务,您只需要提供保存完好的组织或细胞标本,康成的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。

    arraystar公司circrna芯片产品列表


    服务 芯片 规格 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 4,236 mouse circular rnas
    rat circular rna microarray service rat circular rna array v2.0 8×15k 14,145 rat circular rnas
  • ● 全面、系统的circrna收集:circrna研究领域顶尖期刊和权威公共数据库,运用强大的信息筛选系统,挑选出最可靠的circrna。

    ● 高效、精准的circrna检测:剪接位点特异性探针与rnase r预处理双重保障,即便相应线性rna存在也能特异性检测circrna。

    ● 严谨、可靠的spike-in对照:在rna样品中加入一组由ercc (external rna controls consortium)开发的外源性rna,可对rna扩增,标记和芯片杂交过程中产生的实验偏差进行校正,增加样本间结果比较的可靠性。

    ● 详尽、丰富的circrna注释:arraystar mirna靶基因预测软件对circrna上的保守mirna的结合位点进行了预测,并对其中匹配分值较高的位点进行了注释(下图)。此外,我们还提供了与circrna对应的线性mrna信息。根据信息,客户可以很方便地从不同角度研究circrna的作用机制和生物功能。


  • 1. 样品rna抽提
           实验对象为组织样品,取适量(50-100mg)新鲜组织样品或正确保存的组织样品,加1ml的rna抽提试剂trizol(invitrogen),匀浆后抽提rna。
           实验对象为细胞样品,每份样品取1×106~1×107细胞,完全吸去培养液后加1ml的rna抽提试剂trizol,裂解后抽提rna。
    2.  rna质量检测
           使用nanodrop测定rna在分光光度计260nm、280nm和230nm的吸收值,以计算浓度并评估纯度。
           用甲醛电泳试剂进行变性琼脂糖凝胶电泳,检测rna纯度及完整性。
           提供rna qc报告。
    3. rnase r处理
    4. cdna/arna样品合成和标记
    5. 标记效率质量检测
           使用nanodrop检测荧光标记效率,标记效率合格以保证后续芯片实验结果的可靠性。
    6. 芯片杂交
           在标准条件下将标记好的探针和高密度基因组芯片进行杂交。
    7. 图像采集和数据分析
           使用genepix 4000b芯片扫描仪扫描芯片的荧光强度,并将实验结果转换成数字型数据保存,使用配套软件对原始数据进行分析运算。
    8. 提供实验报告:
           • scanning image: cy3荧光扫描图像;
           • scatter plot:散点图,x轴为control信号值,y轴为experiment信号值,表示信号数据总体分布趋势;
           • raw data:探针的扫描荧光信号强度原始数据;
           • normalized data:经过统计学方法标准化后数值;
           • p–value: t-test统计分析显著差异表达的基因,p-value越小,该circrna在两组样本间差异表达越显著;        

           • significant up & down regulated circrna list:显著差异表达的circrna列表包含fold change, p-value。


  • circrna在免疫衰老中的相关研究(comprehensive circular rna profiling reveals that circular rna100783 is involved in chronic cd28-associated cd8( )t cell ageing. immunity & ageing, 2015)

           生物机体免疫系统随着年龄的增长而不断恶化,这个过程被称为免疫衰老(immunosenescence);免疫衰老与系统炎症反应、慢性炎症疾病及癌症发生等密切相关,但目前对引发免疫衰老的机制了解的甚少。该研究通过arraystar circrna芯片筛选发现:在健康成人与免疫力低下的老年人的外周血淋巴细胞中,circrna的表达谱具有明显的差异。在差异表达的circrna中,研究者通过qpcr验证发现hsa_circrna_100783可以明显区分两种不同类型的样本,可以作为免疫衰老的biomarker。研究者接着对circrna可能结合的mirna进行了靶基因预测,推测circrna很可能通过调控磷酸蛋白信号转导途径来影响免疫细胞衰老的过程。

  • → circular rna mto1 acts as the sponge of mir-9 to suppress hepatocellular carcinoma progression.  hepatology. 2017

    → a circular rna binds to and activates akt phosphorylation and nuclear localization reducing apoptosis and enhancing cardiac repair. theranostics. 2017

    → circrna_100290 plays a role in oral cancer by functioning as a sponge of the mir-29 family. oncogene. 2017

    → circular rna_larp4 inhibits cell proliferation and invasion of gastric cancer by sponging mir-424-5p and regulating lats1 expression. molecular cancer. 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

    → changing expression profiles of lncrnas, mrnas, circrnas and mirnas during osteoclastogenesis. scientific reports. 2016

    → circular rna related to the chondrocyte ecm regulates mmp13 expression by functioning as a mir-136 'sponge' in human cartilage degradation. scientific reports. 2016

    → changing expression profiles of lncrnas, mrnas, circrnas and mirnas during osteoclastogenesis. scientific reports. 2016

    → 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 expression profile and potential function of hsa_circrna_101238 in human thoracic aortic dissection. oncotarget. 2017

    → circular rnas signature predicts the early recurrence of stage iii gastric cancer after radical surgery. oncotarget. 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

    → microarray expression profile of circular rnas in peripheral blood mononuclear cells from rheumatoid arthritis patients. cellular physiology and biochemistry. 2017

    → identification of circular rnas as a promising new class of diagnostic biomarkers for human breast cancer. oncotarget. 2017

    → profiling and bioinformatic analysis of circular rna expression regulated by c-myc. oncotarget. 2017

    → circrna expression pattern and circrna-mirna-mrna network in the pathogenesis of nonalcoholic steatohepatitis. oncotarget. 2016

    → comprehensive analysis of differentially expressed profiles of lncrnas and circrnas with associated co-expression and cerna networks in bladder carcinoma.oncotarget. 2016

    → landscape of rnas in human lumbar disc degeneration. oncotarget. 2016

    → circrna_100269 is downregulated in gastric cancer and suppresses tumor cell growth by targeting mir‐630. aging-us. 2017

    → circular rna expression profile of pancreatic ductal adenocarcinoma revealed by microarray. cellular physiology and biochemistry. 2016