Expression Profiling of Cellular Metabolism Using Massively Parallel Sequencing

使用大规模并行测序进行细胞代谢的表达谱分析

• 批准号: 7793135
• 负责人: Helen Haskell Hobbs
• 金额: $ 49.38万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-27 至 2011-05-26
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7793135
• 关键词: Cause of Death; Cholesterol Homeostasis; Communities; DNA Methylation; DNA Sequence; DNA Sequencing Facility; Detection; Diet; Disease; Epigenetic Process; Funding; Future; Gene Expression Profiling; Gene Targeting; Genome; Glean; Goals; Grant; Health; Homeostasis; Human; Laboratories; Ligation; Link; Metabolic; Metabolism; Methylation; Molecular; Molecular Profiling; Obesity; Oligonucleotides; Pattern; Productivity; Recording of previous events; Research; Research Personnel; Research Project Grants; Scientist; Services; Structure; System; Technology; United States National Institutes of Health; Universities; base; cost; direct application; disability; experience; genome-wide; instrument; new technology; next generation; programs; research study; response; transcription factor

DESCRIPTION (provided by applicant): Next-generation sequencing (NGS) provides a powerful new technology to rapidly and effectively interrogate DNA sequence in a simple, comprehensive and cost-effective manner. Probing patterns of whole genome expression, defining the full panoply of genes targeted by transcription factors, and assessing genome-wide methylation status has become feasible and affordable using NGS. The goal of this grant is to establish a high through-put NGS Facility to support ongoing research projects by a thematically-linked cadre of scientists who have a history of implementing new technologies to make high- impact, fundamental discoveries with direct applications to human health. The research focuses on two major causes of death and disability: disorders of cholesterol metabolism and dysregulated fuel homeostasis. Funds are requested to purchase a Sequencing by Oligonucleotide Ligation and Detection (SOLiD) 3 System from Applied Biosystems. The instrument will be operated by the McDermott Center DNA Sequencing Core Laboratory. Initially, the NGS Facility will support investigators funded by two NIH- sponsored programmatic grants: a Program Project (The Molecular Basis of Cholesterol Metabolism) spearheaded by Drs. Michael Brown and Joseph Goldstein that is in its 32nd year of funding, and a new interdisciplinary Roadmap grant, the Taskforce for Obesity Research at UT Southwestern, which brings together 25 independent PI's from 16 different departments. This proposal is structured so that a group of 13 major users with strong scientific programs and a track record of making biologically significant observations will use the SOLiD 3 System to enhance experiments already funded by the NIH. Proposed studies include using gene expression profiling to interrogate the cellular responses to metabolic perturbations, CHiP-sequencing to identify target genes of key metabolic transcription factors discovered by PI's of this application, and DNA methylation profiling to assess the impact of diet on epigenetic programming. Given the scientific productivity of the PI's on this grant, we are optimistic that application of this technology will result in important new discoveries that will directly impact human health. Although the users of the NGS Facility will initially be limited to 13 investigators, the experience gleaned by these scientists will be rapidly disseminated to the entire UT Southwestern research community. We anticipate that both the scope of applications and the spectrum of users will expand in the near future and that the SOLiD System will become an integral part of the McDermott Center DNA Sequencing Core that has provided excellent university-wide service at UT Southwestern for over a decade.

描述（由申请人提供）： 下一代测序（NGS）提供了一种强大的新技术，以简单，全面和成本效益的方式快速有效地询问DNA序列。探测整个基因组表达的模式，定义了由转录因子靶向的基因的完整泛池，并使用NGS评估了全基因组甲基化状态的可行和负担得起。这笔赠款的目的是建立一个高的贯穿NGS设施，以支持由主题链接的科学家干部进行的正在进行的研究项目，他们有实施新技术的历史，以对人类健康直接适用于人类健康。该研究侧重于死亡和残疾的两个主要原因：胆固醇代谢和燃料稳态失调的疾病。 要求资金从Applied Biosystems购买寡核苷酸连接和检测（实心）3系统的测序。该仪器将由McDermott Center DNA测序核心实验室操作。最初，NGS设施将支持由两名NIH赞助的计划赠款资助的研究人员：一个由DRS率领的计划项目（胆固醇代谢的分子基础）。迈克尔·布朗（Michael Brown）和约瑟夫·戈德斯坦（Joseph Goldstein）是其第32年的资助，以及新的跨学科路线图格兰特（UT）的新跨学科路线图格兰特（UT）的肥胖工作组，该工作人员从16个不同部门汇集了25个独立PI。 该提案的结构化，以便由13个具有强大科学计划的主要用户组成的组，并具有使生物学上重要的观察结果的记录将使用Solid 3系统来增强NIH已经资助的实验。拟议的研究包括使用基因表达分析来询问对代谢扰动的细胞反应，CHIP测序以鉴定PI在该应用中发现的关键代谢转录因子的靶基因，以及该应用的DNA甲基化分析以评估饮食对表观遗传程序的影响。鉴于PI在这笔赠款中的科学生产力，我们乐观地认为，该技术的应用将导致重要的新发现，这将直接影响人类健康。尽管NGS设施的使用者最初将仅限于13位调查人员，但这些科学家收集的经验将迅速传播到整个UT西南研究社区。我们预计，应用程序的范围和用户的范围都将在不久的将来扩展，并且固体系统将成为McDermott Center DNA测序核心不可或缺的一部分，该核心已在UT Southwestern提供了出色的服务已有十多年了。