Creation of mouse strains for the study of hematopoiesis

用于造血研究的小鼠品系的创建

• 批准号: 6757294
• 负责人: Archibald S. Perkins
• 金额: $ 16.35万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-08-01 至 2005-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6757294
• 关键词: Drosophilidae; biotechnology; blood cell count; cell differentiation; cell growth regulation; cytogenetics; ecdysone; gene expression; gene mutation; genetic library; genetic screening; genetic strain; genetically modified animals; green fluorescent proteins; hematopoiesis; hematopoietic stem cells; high throughput technology; hormone regulation /control mechanism; laboratory mouse; luciferin monooxygenase; melanocyte stimulating hormone; nitrosourea; pluripotent stem cells; regulatory gene; reporter genes; technology /technique development; tissue /cell culture

DESCRIPTION (provided by applicant): The availability of DNA sequence for the human and mouse has heralded a new era in biology. These data are yielding structural details for all human and mouse genes. Our ability to interpret and use these data is dependent on our ability to assess gene function within a whole organism, which is typically done through the study of gene mutations. For the study of human disease, especially those of the blood, the mouse constitutes a premier model. In the mouse, it is possible to induce random point mutations in the germline using ethyl nitrosourea (ENU). ENU mutagenesis of mice followed by screens for specific phenotypes allows for the isolation of strains bearing single gene mutations that cause deviation from normal function. This, coupled with robust approaches to mutant gene identification, allows the identification of genes that regulate complex biological processes. Because the mutations are single base pair changes, the alleles generated are very often partial loss of function alleles, and thus no redundant allele series are possible, which can give a wealth of information about gene function. In addition, gain-of-function alleles are also possible. The power of ENU mutagenesis is entirely dependent on the sensitivity and specificity of the phenotypic screen. We propose the development of mouse strains that will allow one to screen for mutants that are altered in particular blood cell compartments. We are interested in the homeostatic mechanisms that regulate hematopoietic stem cell number and commitment to differentiation. This is a complex process that is under genetic control. Indeed, it has been possible to identify variability in stem cell number among mouse strains and to map several influential genes, but as of yet, no specific genes that regulate stem cell number have been identified. The approaches described thus far are hampered by major problems including the laborious nature of screens for blood cell number and function. We intend to generate strains of reporter mice that will allow the enumeration of a specific blood cell lineage compartment through analysis of mouse serum. The establishment of such strains will allow one to screen for mutants that have abnormal hematopoiesis.

描述（由申请人提供）：人类和小鼠DNA序列的可用性预示着生物学的新时代。这些数据提供了所有人类和小鼠基因的结构细节。我们解释和使用这些数据的能力取决于我们在整个生物体中评估基因功能的能力，这通常是通过研究基因突变来完成的。对于人类疾病的研究，尤其是血液疾病的研究，老鼠是一个首要的模型。在小鼠中，使用乙基亚硝基脲（ENU）可以在种系中诱导随机点突变。对小鼠进行ENU诱变，然后对特定表型进行筛选，从而分离出导致正常功能偏离的单基因突变菌株。这与突变基因鉴定的稳健方法相结合，允许鉴定调节复杂生物过程的基因。由于突变是单碱基对的改变，因此产生的等位基因往往是部分功能缺失的等位基因，因此不可能存在冗余的等位基因序列，这可以提供丰富的基因功能信息。此外，获得功能等位基因也是可能的。ENU诱变的能力完全取决于表型筛选的敏感性和特异性。我们建议开发小鼠品系，使人们能够筛选在特定血细胞区室中发生改变的突变体。我们对调节造血干细胞数量和分化承诺的稳态机制感兴趣。这是一个受基因控制的复杂过程。事实上，已经有可能确定小鼠品系之间干细胞数量的可变性，并绘制出几个有影响的基因，但到目前为止，还没有确定调节干细胞数量的特定基因。迄今为止所描述的方法受到主要问题的阻碍，包括筛选血细胞数量和功能的费力性。我们打算产生报告小鼠的菌株，将允许枚举一个特定的血细胞谱系室通过小鼠血清分析。建立这样的菌株将允许人们筛选具有异常造血功能的突变体。