Consortium for large-scale production and cryopreservation of knockout mice

基因敲除小鼠大规模生产和冷冻保存联盟

• 批准号: 8189478
• 负责人: MONICA J. JUSTICE
• 金额: $ 413.3万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8189478
• 关键词: Consortium; large; scale; production; cryopreservation

DESCRIPTION (provided by applicant): Annotation of the reference human genome has identified approximately 20,000 protein coding genes as well as 3,000 non-coding RNAs. Together these genes orchestrate the development of the organism, supporting all aspects of the function of cells, tissues, organ systems as well physiology and behavior. The l000 genomes project has revealed extraordinary levels of diversity in human genomes, yet for most genes neither the function of normal version nor the disease consequence of loss-of-function variants is known. The mouse provides a route to understand the function of genes and their variants. Mice share developmental, physiological, anatomical and metabolic parallels with humans, which are evident in healthy as well as diseased states. These reflect similarities of the genes in both species. Mutant mice generated using ES cell technology are a sensitive biological assay system from which a deep understanding of function can be gleaned and they also provide a long lasting biological resource for further study. The objective of this proposal is to generate mutant mice from a resource of ES cells with conditionally targeted, lacZ-tagged alleles generated under previous NIH (KOMP) and EU (EUCOMM) funded programs. We and others will use these mice to discover the function of genes. We are proposing to carry out this work at scale and have formed a consortium of three Institutes (Baylor College of Medicine, the Sanger Institute and MRC Harwell) to work together as equal partners to achieve this objective. We will generate mice corresponding to 1500 mutant genes from ES cells distributed by the Sanger Institute. We will characterize the adult and embryonic expression pattern of the each targeted gene and by breeding determine the requirement of each for embryonic development and fertility, if any. All of the alleles will be cryo-preserved and placed in a repository and all of the data will be deposited in a centralized data coordination center to support further studies. RELEVANCE: Most of the 20,000 genes in a typical person are normal but we also have several hundred broken ones. While some broken genes can cause severe disease such as cystic fibrosis or cancer, others have little or no consequence, or function only under stress. Currently we have some understanding of the function of just one third of human genes. If we are to fully understand human health and disease we must expand knowledge of gene function to all of our genes.

描述（由申请人提供）：参考人类基因组的注释已确定约20,000个蛋白质编码基因以及3,000个非编码RNA。这些基因共同策划了生物体的发展，支持细胞，组织，器官系统以及生理学和行为的各个方面。 L000基因组项目揭示了人类基因组的多样性水平，但是对于大多数基因而言，既不是正常版本的功能，也不是功能丧失变体的疾病后果。小鼠提供了了解基因及其变体功能的途径。小鼠与人类具有发育，生理，解剖学和代谢的相似之处，在健康和患病状态中都很明显。这些反映了两个物种中基因的相似性。使用ES细胞技术产生的突变小鼠是一种敏感的生物学测定系统，可以从中收集对功能的深刻理解，并且还为进一步研究提供了持久的生物学资源。该提案的目的是从ES细胞资源中产生突变小鼠，并具有有条件的靶向LACZ标记等位基因（KOMP）和欧盟（EUCOM）（Eucomm）资助的程序。我们和其他人将使用这些小鼠发现基因的功能。我们建议按大规模开展这项工作，并组成了三个学院（贝勒医学院，桑格学院和MRC Harwell）的财团，作为平等合作伙伴共同努力，以实现这一目标。我们将产生与Sanger Institute分布的ES细胞1500个突变基因相对应的小鼠。我们将表征每个靶向基因的成人和胚胎表达模式，并通过育种来确定每个靶向基因对胚胎发育和生育的需求（如果有的话）。所有等位基因将是冷冻保存并放置在存储库中的，所有数据将存放在集中式数据协调中心中，以支持进一步的研究。 相关性：一个典型人的20,000个基因中的大多数是正常的，但我们也有数百个破碎的基因。尽管某些破裂的基因会引起严重的疾病，例如囊性纤维化或癌症，但其他基因几乎没有或根本没有压力，或者仅在压力下起作用。目前，我们对仅三分之一的人类基因的功能有所了解。如果我们要充分了解人类健康和疾病，则必须将对基因功能的知识扩展到我们的所有基因。

项目成果

Rapid conversion of EUCOMM/KOMP-CSD alleles in mouse embryos using a cell-permeable Cre recombinase.

• DOI: 10.1007/s11248-013-9764-x
• 发表时间: 2014-02
• 期刊: TRANSGENIC RESEARCH
• 影响因子: 3
• 作者: Ryder, Edward;Doe, Brendan;Gleeson, Diane;Houghton, Richard;Dalvi, Priya;Grau, Evelyn;Habib, Bishoy;Miklejewska, Evelina;Newman, Stuart;Sethi, Debarati;Sinclair, Caroline;Vyas, Sapna;Wardle-Jones, Hannah;Bottomley, Joanna;Bussell, James;Galli, Antonella;Salisbury, Jennifer;Ramirez-Solis, Ramiro
• 通讯作者: Ramirez-Solis, Ramiro