Functionally Validated Lentiviral siRNA Libraries

功能验证的慢病毒 siRNA 文库

基本信息

批准号：
7275220
负责人：
ALEX CHENCHIK
金额：
$ 28.94万
依托单位：
CELLECTA, INC.
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-07-06 至 2009-07-05
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7275220
关键词：
Address Animal Model Antineoplastic Agents Apoptosis Attention Biological Biological Assay Biological Models Biology Cell Line Cell Separation Cell model Cells Chemicals Clinical Collaborations Collection Compatible Cultured Cells Custom Cytochrome P450 Development Disease Dissection Drug Delivery Systems Eligibility Determination Exons Facility Construction Funding Category Fred Hutchinson Cancer Research Center Funding Gene Family Gene Targeting Generations Genes Genetic Screening Genome Genomics Goals Green Fluorescent Proteins Growth Human Human Genome Project Individual Informatics Ion Channel Knowledge Laboratories Lentivirus Vector Libraries Link Malignant Epithelial Cell Mammalian Cell Messenger RNA Modeling Molecular Mus Normal Cell Nuclear Hormone Receptors Pathology Pathway interactions Performance Pharmaceutical Preparations Pharmacologic Substance Phase Phase I Clinical Trials Phenotype Primary carcinoma of the liver cells Process Protein Kinase Proteins Protocols documentation Publishing Puromycin RNA Interference Range Reagent Reporter Research Research Institute Research Personnel Resistance Resources Screening procedure Services Signal Pathway Small Business Funding Mechanisms Small Business Innovation Research Grant Small Interfering RNA Stem cells System Technology Tetracycline Tetracyclines Therapeutic Intervention Translational Research Validation base cancer cell cellular transduction concept cost design design and construction desire drug discovery experience gene function high throughput technology human disease improved loss of function mouse genome novel novel therapeutics particle programs receptor research study small hairpin RNA small molecule structural biology success therapy development tool

Address Animal Model Antineoplastic Agents Apoptosis Attention Biological Biological Assay Biological Models Biology Cell Line Cell Separation Cell model Cells Chemicals Clinical Collaborations Collection Compatible Cultured Cells Custom Cytochrome P450 Development Disease Dissection Drug Delivery Systems Eligibility Determination Exons Facility Construction Funding Category Fred Hutchinson Cancer Research Center Funding Gene Family Gene Targeting Generations Genes Genetic Screening Genome Genomics Goals Green Fluorescent Proteins Growth Human Human Genome Project Individual Informatics Ion Channel Knowledge Laboratories Lentivirus Vector Libraries Link Malignant Epithelial Cell Mammalian Cell Messenger RNA Modeling Molecular Mus Normal Cell Nuclear Hormone Receptors Pathology Pathway interactions Performance Pharmaceutical Preparations Pharmacologic Substance Phase Phase I Clinical Trials Phenotype Primary carcinoma of the liver cells Process Protein Kinase Proteins Protocols documentation Publishing Puromycin RNA Interference Range Reagent Reporter Research Research Institute Research Personnel Resistance Resources Screening procedure Services Signal Pathway Small Business Funding Mechanisms Small Business Innovation Research Grant Small Interfering RNA Stem cells System Technology Tetracycline Tetracyclines Therapeutic Intervention Translational Research Validation base cancer cell cellular transduction concept cost design design and construction desire drug discovery experience gene function high throughput technology human disease improved loss of function mouse genome novel novel therapeutics particle programs receptor research study small hairpin RNA small molecule structural biology success therapy development tool

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项目摘要

DESCRIPTION (provided by applicant): Despite the recent completion of the human genome project, an ostensibly more difficult post-genomic challenge will be the functional annotation of all human genes and integration of this information into an operational cell-based model. Unfortunately, this is at present challenging, primarily due to the absence of suitable toolsets to rapidly delineate gene function en masse. RNA interference (RNAi) has proven to be an extremely potent and versatile tool to specifically reduce expression of targeted genes, allowing for loss-of- function genetic screens in mammalian cells. Despite these successes, high-throughput (HT) RNAi screening is technically challenging and significant limitations in the technology exist. To address these issues, and to expand on previous program funding, we have developed a novel experimental platform to construct functionally validated (FV) shRNA libraries. Under Phase I funding, we initially propose to develop a FV human shRNA library in biosafe lentiviral vectors targeting 5000 pharmaceutically tractable gene families that are amenable to small molecule drug discovery research. In collaboration with our biology consultants at the Fred Hutchinson Cancer Research Center (FHCRC) and the Scripps Research Institute (TSRI), we will subsequently validate this library in a simple cell culture-based model, to identify genes responsible for the growth limiting actions of TGF-¿ in cancer cells. In Phase II, we will extend this program to include the development of genome-wide functionally validated human and mouse siRNA libraries. We propose to use our novel HT RNAi resource to delineate the processes which underlie deregulated proliferation in cancer cells, and to apply this knowledge into translational research programs. For example, we will use our newly developed FV siRNA library technology to identify synthetic lethal interactions in cancer cells. Genetic screens with FV siRNA libraries have the potential to greatly simplify gene manipulation and discovery for many biomedical applications, such as validation of gene function, probing interactions between genes, and the establishment of animal models. Moreover, we envisage a major impact on the molecular dissection of human disease mechanisms. For example, these reagents harbor considerable promise to identify new targets for therapeutic intervention, and the development of increasingly relevant paradigms for drug discovery. As a result, we foresee that these toolsets will significantly improve the efficiency, economy and ease of performing HT RNAi screens, and will provide basic researchers with preferred, cost-effective alternatives to existing commercially available reagents. The ultimate goal of the proposed project is to develop and make commercially available a new powerful research tool: a set of functionally validated genome-wide human and mouse lentiviral siRNA libraries designed for high-throughput discovery of novel drug targets. We propose to apply this powerful technology to identify novel anti- cancer drug targets. The developed technologies will significantly improve the efficiency of translational researches related to molecular dissection of diverse human disease mechanisms, development of new pharmaceuticals and therefore, have major implications for improving drug discovery research.

描述（由适用提供）：尽管人类基因组项目最近完成，但表面上更困难的基因组挑战将是所有人类基因的功能注释以及将该信息整合到基于操作细胞的模型中。不幸的是，这是当前的挑战，主要是由于没有合适的工具集来快速描绘基因功能。 RNA干扰（RNAi）已被证明是一种极度潜在的和多功能的工具，可特别减少靶向基因的表达，从而允许哺乳动物细胞中功能丧失的遗传筛选。尽管取得了这些成功，但高通量（HT）RNAi筛查在技术上是挑战，并且在技术中存在重大局限性。为了解决这些问题，为了扩展以前的计划资助，我们开发了一个新颖的实验平台来构建经验验证的SHRNA库。在第一阶段的资金下，我们最初建议在生物保护慢病毒载体中开发FV人类shRNA文库，其针对5000种可容纳针对小分子药物发现研究的药物可牵引基因家族。在弗雷德·哈钦森癌症研究中心（FHCRC）和Scripps研究所（TSRI）的生物学顾问中，我们随后将以简单的基于细胞培养的模型来验证该图书馆，以识别负责癌细胞中TGF-限制性作用的基因。在第二阶段，我们将扩展该程序，以包括全基因组功能验证的人和小鼠siRNA库的发展。我们建议利用我们的新型HT RNAi资源来描述癌细胞中放松癌细胞增殖的过程，并将这些知识应用于翻译的研究计划。例如，我们将使用新开发的FV SiRNA库技术来鉴定癌细胞中的合成致命相互作用。具有FV SIRNA库的遗传筛选有可能大大简化许多生物医学应用的基因操纵和发现，例如基因功能的验证，基因之间的相互作用以及动物模型的建立。此外，我们设想对人类疾病机制的分子解剖产生重大影响。例如，这些试剂具有巨大的承诺，可以识别治疗干预的新目标，并开发了越来越多的相关药物发现范式。结果，我们预见到这些工具集将显着提高执行HT RNAi屏幕的效率，经济性和易度性，并将为基础研究人员提供现有市售试剂的首选，具有成本效益的替代方法。拟议项目的最终目标是开发并使商业上可用的新功能研究工具：一组由功能验证的全基因组和小鼠慢病毒siRNA库库，旨在高通量发现新型药物靶标。我们建议应用这项强大的技术来识别新型的抗癌药物靶标。开发的技术将显着提高与分子解剖有关的人类疾病机制，新药的开发，因此对改善药物发现研究具有重要意义。