LncRNA全国讲座.ppt

单击此处编辑母版标题样式,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,*,*,单击此处编辑母版标题样式,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,*,*,单击此处编辑母版标题样式,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,*,*,长链非编码,RNA,研究思路,“Learner&Nontrivial&Confidence”-RNA,施沫,全国技术中心 技术支持经理,上海其明信息技术有限公司,1,L,earner,“You start projects and you have no idea what to expect.Its that kind of challenge that I enjoy very much as a scientist.”,2,Nontrivial,Circ Res,2012,3,Confidence,Therapeutic approach,In clinical trials,Approved therapy,Enzyme replacement therapy,YES,YES,Gene therapy,YES,YES,Nuclear receptor ligands,YES,YES,Chromatin or DNA modulators,YES,YES,Artificial transcription factors,YES,NO,MicroRNA modulators,YES,NO,Cis,-acting lncRNA modulators,NO,NO,Trans,-acting lncRNA modulators,NO,NO,4,非,RNA,利用医薬開発,Genome Network,Protein 2,Protein 1,Protein 3,Protein 4,Protein 5,Protein 5,P,Gene 2,Protein X,Gene 10,ncRNA 3,ncRNA 4,Gene 5,Gene 6,Drug,Gene 7,Protein 10,Protein 12,Gene 3,Protein 7,Protein 7,Protein 8,Protein 9,Gene 1,Protein 11,Protein 13,ncRNA 1,ncRNA 6,Gene 4,Gene 11,Gene 9,ncRNA 2,Gene 8,Disease,New Target of Medicine,Mutsumi Kanamori-Katayama Ph.D.,Yoshihide Hayashizaki M.D.,Ph.D.,RIKEN Genome Sciences Center Genome Exploration Research Group,5,物种间数量变化趋势,6,7,Ryan et.al,BioEssays 29:288299(2007),8,LncRNA,200nt,结构类似,mRNA,具有外显子,/,内含子,低蛋白编码潜力,Non Coding RNA,分类,9,LncRNA,来源,10,LncRNA,表达特异性,Cabili MN.Integrative annotation of human large intergenic noncoding RNAs reveals global properties and specific subclasses.Genes Dev.2011 Sep 15;25(18):1915-27,11,lncRNA,调控方式案例,12,ncRNA,调控,13,lncRNA,反式与顺式调控,14,lncRNA,的反式调控方式,ncRNA PANDA,募集转录因子,NF-YA,，启动细胞凋亡通路,15,反式调控,Cell 2011,ncRNA-and Pc2 Methylation-Dependent Gene Relocation between Nuclear Structures Mediates Gene Activation Programs,16,l,ncRNA,的顺式调控方式,稳定,RNA,Nature Medicine 2010,Expression of a,noncoding RNA,is elevated in Alzheimers disease and drives rapid feed-forward regulation of-secretase expression,早发老年性痴呆病中,BACE1-AS,高表达导致,BACE1 mRNA,稳定性增高，蛋白积聚，体现了,ncRNA,的毒性效应,BACE1,BACE1-AS,17,拷贝数,染色体定位,表达,18,Science,案例分析,19,其明成功案例,IF:31.027,20,所涉及的实验技术,21,实验材料,22,实验结果,芯片结果聚类图,测序结果散点图,back,23,Northern blot,lncRNA-DC,在免疫细胞中的表达情况,lncRNA-DC,在,DC/mono,细胞群中的表达情况,back,24,ChIP-Seq,与,ChIP-PCR,结果,back,25,荧光素酶报告分析，验证转录因子结合区域,荧光素酶报告实验,back,26,LNC-DC,行驶的细胞功能研究,干扰结果,27,过表达结果,back,28,小鼠干扰模型,抑制,DC,细胞分化,back,29,通过,STAT3,的,RIP,实验验证,RIP&RNA Pull down,Lnc-DC,与,STAT3,相互结合,back,30,进一步验证,Pull down,和,RIP,的结果,细胞定位实验,back,31,结果表明，,lnc-DC,的,3,末端序列（,265-417,）与,STAT3,的,C,端结合（,583-770,）。,STAT3,的活化依赖,C,端,Tyr,705,的磷酸化。,截断突变实验,back,32,研究结果表明，,S3I-201,处理的细胞系与,RNAi,干扰后的细胞系在基因表达差异方面非常相似。,药物干预,33,思路,高通量检测,生物信息分析,表达、定位验证,上游,验证,功能,验证,下游,验证,34,为何,选择,HTA2.0,联合检测,35,36,HTA2.0,优势,37,可变剪切,生物信息学分析,38,HTA2.0,可变剪切变得如此简单,Affymetrix Transcriptome Analysis Console 2.0(TAC 2.0)Software,39,引物设计,40,与,PCR,验证的吻合度,41,案例,IF:10.143,42,案例,IF:4,IF:11.139,43,基因组位置分析,生物信息学分析,44,lncRNA,基因组位置关系分析,上游,下游,反义链,正义链,45,LncRNA,基因组结果应用,46,非编码,RNA,调控,生物信息学分析,47,共表达网络,长链非编码,RNA,与编码基因互作,48,Hepatology 2011,LncRNA-mRNA,相互作用网络应用,49,通过,网络,找到,调控,lncRNA,最多的,转录因子,同时找到能被不同转录因子调控最多的,lncRNA,使用,德国,Biobase,数据库商业版数据库,lncRNA,反式分析,TF-lncRNA-network,50,新视角,反馈环路分析,Affymetrix Promoter1.0R Array,Affymetrix HTA2.0,51,新视角,前馈环路分析,蓝色线表示两点之间的关系需要计算,绿色线表示两点之间的关系通过转录因子的预测得到,52,competing endogenous RNAs(ceRNAs),分析,53,54,案例,IF:38.597,55,56,深灰色：激活,浅灰色：抑制,57,案例,IF:7.357,58,lncRNA,相关芯片与分析服务,59,其明生物,值得信赖的合作伙伴,60,上海其明介绍,61,CPU,256,颗,CPU,保证繁重运算顺利进行,不会死机,内存,2048G,可同时运行,256,个样本,的,数据分析,硬盘,2.3P,可存储,2.310,7,G,数据,硬件平台,62,软件平台,63,上海其明优势,64,施沫,全国统一服务电话：,40030-40060,公司电话：,021-62228901-211,手机：,13761936890,Email:,shimo,Thank You,For Your Attention,65,