Comparative Functional Genomics of Longevity Assurance

描述（由申请方提供）：我们建议通过一组重组近交（RI）线虫和不同预期寿命的小鼠品系以及长寿突变线虫品系的全局基因表达谱来表征长寿保证基因（LAG）。我们的实验设计-这是从其比较性质的权力-旨在确定这些基因（a）的转录水平始终与跨RI线和突变体与野生型动物的寿命长，和（B），这种协会被发现独立于多个物种。这种实验方法将需要在生物学、遗传学、基因组学和统计学方面的协调努力，聚集的团队成员反映了这种多样性。我们的具体目标是： 1.定量描述两种线虫12个RI系的寿命。 将确认C的六条现有RI线的平均和最大寿命。elegans和6个新分离的C. briggsae; 2.为了验证利用全基因组比较表达分析技术对水稻RI株系和突变株进行基因组比较表达分析的假设。elegans将识别LAG候选人。利用全基因组芯片分析了6个RI株系和2个突变株的基因表达。elegans将在固定的时间和生理年龄进行。我们将使用高灵敏度的Q-PCR分析来验证从微阵列分析中挑选的候选基因的表达水平; 3.确定特定目标2中表征的候选基因中的哪些是必需的LAG。利用RNAi技术来操纵特定目标2中发现的候选基因在C. elegans将允许分配给这些基因的相对权重; 4.为鉴定重组自交系中的LAG候选基因。briggsae通过转录谱。这一具体目标将（a）使我们能够表征“私人”与“公共”机制，以增加预期寿命，（B）允许一组独立的数据来支持（或反驳）这些数据产生的RNAi在具体目标3。 5.将分析方法扩展到小鼠RI系。在第四年和第五年，我们将应用上述定义的功能基因组方法，以及前三年改进的统计方法，分析具有不同寿命的小鼠RI系。

DESCRIPTION (provided by applicant): We propose to characterize longevity assurance genes (LAGs) through global gene expression profiling of a panel of recombinant inbred (RI) nematode and mouse strains of varying life expectancy, as well as long-lived mutant nematode strains. Our experimental design--which derives power from its comparative nature--aims to identify those genes (a) for which transcriptional level is consistently associated with long lifespan across RI lines and in mutant versus wild-type animals, and (b) for which such associations are found independently in multiple species. This experimental approach will require coordinated efforts in organismal biology, genetics, genomics, and statistics, and the team members assembled reflect this diversity. Our specific aims are: 1. To quantitatively characterize the lifespans of twelve nematode RI lines in two species. Mean and maximum lifespans will be confirmed for six existing RI lines of C. elegans and six newly-isolated RI lines of C. briggsae; 2. To test the hypothesis that whole-genome comparative expression analysis of RI lines and mutant strains of C. elegans will identify LAG candidates. Whole-genome microarray analysis of gene expression in six RI lines and two mutant strains of C. elegans will be carried out, at fixed chronological and physiological ages. We will employ highly sensitive Q-PCR analysis to validate the expression levels of candidate genes picked from microarray analysis; 3. To determine which of the candidate genes characterized in Specific Aim 2 are necessary LAGs. The use of RNAi technology to manipulate the expression of a subset of candidate genes discovered in Specific Aim 2 in C. elegans will permit a relative weight to be assigned to these genes; 4. To identify LAG candidates in recombinant inbred lines of C. briggsae via transcriptional profiling. This specific aim will (a) allow us to characterize "private" versus "public" mechanisms for increasing life expectancy, and (b) permit an independent set of data to support (or contradict) those data generated with the use of RNAi in Specific Aim 3. 5. To extend the analytical approach to mouse RI lines. In years four and five we will apply the functional genomic approach defined above, as well as statistical methods refined during the first three years, to the analysis of mouse RI lines with differential lifespans.