Creating Comprehensive Maps of Worm and Fly Transcription Factor Binding Sites
创建蠕虫和苍蝇转录因子结合位点的综合图谱
基本信息
- 批准号:9526117
- 负责人:
- 金额:$ 91.57万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2013
- 资助国家:美国
- 起止时间:2013-09-20 至 2018-05-31
- 项目状态:已结题
- 来源:
- 关键词:Amino Acid SequenceAnimal ModelAnimalsAntibodiesBacterial Artificial ChromosomesBedsBindingBinding SitesBioinformaticsBiological TestingBiologyCaenorhabditis elegansCodeCommunitiesDataData QualityData SetDatabasesDevelopmentDiseaseDrosophila genusDrosophila melanogasterElementsFoundationsFundingGene ExpressionGene Expression ProfilingGene FusionGene TargetingGene Transfer TechniquesGenesGenomeGenomicsGrantHealthHumanLife Cycle StagesLinkMapsMusNational Human Genome Research InstituteNematodaOrganismPatternPhasePrincipal InvestigatorProcessProductionPropertyPublicationsRNA InterferenceRegulatory ElementResearchScienceSiteStandardizationStudy modelsTestingTimeTranscriptUnited States National Institutes of HealthValidationWorkbasecomputerized data processingenhanced green fluorescent proteinexperimental studyflygenome sequencinghuman diseaseimprovedin vivoinsightknock-downmembermodel organisms databasestranscription factortranscriptometranscriptome sequencing
项目摘要
DESCRIPTION (provided by applicant): Our project seeks to identify the regulatory elements recognized by the vast majority of transcription factors (TFs) in the fruit fly Drosophila melanogaster and the nematode Caenorhabditis elegans. In the initial modENCODE project, an experimental pipeline was developed and applied to -100 TFs in each organism. In this intervening year, we expect to capture data for another 75-100 factors. The present project builds on the advances made by the groups in the initial phase and also combines the production pipelines to increase efficiency and to realize economies of scale. With these improvements, we will generate data sets for another 400 factors from each organism, which when combined with previous work will represent the bulk of all transcription factors in these key model organisms. For both organisms, the overall strategy tags transcription factor genes by fusion with an enhanced Green Fluorescent Protein (eGFP) sequence through recombineering of large insert clones, and introducing the tagged genes into the genome by transgenesis. ChlP-seq using a high quality anti-GFP antibody is performed on the developmental stage(s) with maximal GFP expression, as guided by available RNA-seq expression data. The aligned sequence reads are analyzed to identify candidate binding sites and likely target genes. We will prioritize TFs with human homologs to maximize the broader utility of the data. We will also perform RNAi of 125 TFs in each organism, followed by RNA-seq, to validate called peaks and their assigned target genes. Finally, we will integrate the information for the different data setsto construct regulatory networks implied by the TF binding site data. We will coordinate with ENCODE projects on human TFs, and our data will provide key in vivo and developmental regulatory information that will be essential to delineate both fundamentally conserved as well as human-specific properties of TFs. RELEVANCE (See instructions): Insights from the study of the model organisms Drosophila and C. elegans provide the basis for broad understanding of fundamental processes of animal biology. Because many of their genes have clear relatives in humans, these studies have also led directly to improved understanding of human diseases and in some cases to therapies. Similarly, creating a comprehensive understanding of transcription factor binding sites and building regulatory networks in these key model organisms will create the foundation for understanding human regulatory networks both in health and disease.
描述(由申请人提供):我们的项目旨在鉴定果蝇和线虫中绝大多数转录因子(TF)识别的调控元件。在最初的modENCODE项目中,开发了实验管道并应用于每种生物体中的约100个TF。在这一年中,我们预计将捕获另外75-100个因素的数据。目前的项目以各集团在初期阶段取得的进展为基础,并结合生产管道,以提高效率和实现规模经济。通过这些改进,我们将从每个生物体中生成另外400个因子的数据集,这些数据集与以前的工作相结合时,将代表这些关键模型生物体中所有转录因子的大部分。对于这两种生物体,总体策略通过与增强的绿色荧光蛋白(eGFP)序列融合来标记转录因子基因,所述融合通过大插入克隆的重组工程进行,并通过转基因将标记的基因引入基因组中。使用高质量抗GFP抗体的ChIP-seq在具有最大GFP表达的发育阶段进行,如由可用的RNA-seq表达数据指导的。分析比对的序列读段以鉴定候选结合位点和可能的靶基因。我们将优先考虑与人类同源的TF,以最大限度地扩大数据的效用。我们还将在每个生物体中对125个TF进行RNAi,然后进行RNA-seq,以验证所谓的峰及其指定的靶基因。最后,我们将整合不同数据集的信息,构建TF结合位点数据所隐含的调控网络。我们将与ENCODE项目对人类转铁蛋白进行协调,我们的数据将提供关键的体内和发育调控信息,这些信息对于描述转铁蛋白的基本保守特性和人类特异性特性至关重要。相关性(见说明):从研究模式生物果蝇和C。线虫为广泛理解动物生物学的基本过程提供了基础。由于它们的许多基因在人类中有明确的亲属,这些研究也直接导致了对人类疾病的理解,并在某些情况下导致了治疗。同样,在这些关键模式生物中建立对转录因子结合位点的全面理解和建立调控网络将为理解人类健康和疾病的调控网络奠定基础。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
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ROBERT H WATERSTON其他文献
ROBERT H WATERSTON的其他文献
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{{ truncateString('ROBERT H WATERSTON', 18)}}的其他基金
Creating Comprehensive Maps of Worm and Fly Transcription Factor Binding Sites
创建蠕虫和苍蝇转录因子结合位点的综合图谱
- 批准号:
8904695 - 财政年份:2013
- 资助金额:
$ 91.57万 - 项目类别:
Creating Comprehensive Maps of Worm and Fly Transcription Factor Binding Sites
创建蠕虫和苍蝇转录因子结合位点的综合图谱
- 批准号:
8737930 - 财政年份:2013
- 资助金额:
$ 91.57万 - 项目类别:
High throughput methods for Synthetic Genetic Array Analysis in C. elegans
线虫合成基因阵列分析的高通量方法
- 批准号:
8490069 - 财政年份:2013
- 资助金额:
$ 91.57万 - 项目类别:
Creating Comprehensive Maps of Worm and Fly Transcription Factor Binding Sites
创建蠕虫和苍蝇转录因子结合位点的综合图谱
- 批准号:
8566279 - 财政年份:2013
- 资助金额:
$ 91.57万 - 项目类别:
Creating Comprehensive Maps of Worm and Fly Transcription Factor Binding Sites
创建蠕虫和苍蝇转录因子结合位点的综合图谱
- 批准号:
9119534 - 财政年份:2013
- 资助金额:
$ 91.57万 - 项目类别:
High throughput methods for Synthetic Genetic Array Analysis in C. elegans
线虫合成基因阵列分析的高通量方法
- 批准号:
8653976 - 财政年份:2013
- 资助金额:
$ 91.57万 - 项目类别:
Comprehensive Identification of Worm and Fly Transcription Factor Binding Sites
蠕虫和苍蝇转录因子结合位点的综合鉴定
- 批准号:
8402441 - 财政年份:2012
- 资助金额:
$ 91.57万 - 项目类别:
USING MACHINE LEARNING TO SPEED UP MANUAL IMAGE ANNOTATION
使用机器学习加速手动图像注释
- 批准号:
8171453 - 财政年份:2010
- 资助金额:
$ 91.57万 - 项目类别:
Global Identification of transcribed elements in the C. elegans genome
线虫基因组中转录元件的整体鉴定
- 批准号:
7923469 - 财政年份:2009
- 资助金额:
$ 91.57万 - 项目类别:
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