Positional cloning of obesity genes from congenic mice

同系小鼠肥胖基因的定位克隆

• 批准号: 7099830
• 负责人: CRAIG H WARDEN
• 金额: $ 33.3万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-15 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7099830
• 关键词: RNA; aging; bioenergetics; diet; gene expression; genetic regulatory element; genetic strain; genetically modified animals; genotype; inbreeding; laboratory mouse; messenger RNA; microRNAs; microarray technology; nucleic acid sequence; nutrient intake activity; obesity; phenotype; polymerase chain reaction; positional cloning; sex

DESCRIPTION (provided by applicant): Positional cloning - the identification of obesity genes by co-incidence of mapped obesity traits and genes to a specific chromosomal region, provides a powerful method to discover novel obesity genes and to make novel insights into the mechanisms causing obesity. Mice have been used for the first identification of more human obesity genes than any other model. The goal of this proposal is to use positional cloning methods to identify obesity genes in mouse congenic strains. Congenic mouse strains are identical to a background strain except for a chromosomal region from a donor strain. Phenotype differences between congenic and background are due to alleles of the donor strain with functional effects different from the background strain. The two laboratories contributing to this proposal have identified several mouse congenic strains with statistically significant phenotypes for obesity on mouse chromosome 2 in a region homologous to human chromosome 20, where several obesity quantitative trait loci have been mapped. Our general hypothesis is that one or more genes or transcripts in the congenic donor regions influence obesity. Our specific working hypothesis is that genes/transcripts underlying obesity can be identified for most congenics. We propose seven Specific Aims. We will: determine diet, sex and age effects on obesity in the founding congenics (Aim 1), identify minimal chromosomal loci containing obesity genes by breeding congenics with ever smaller donor regions that retain obesity phenotypes (Aim 2), find differentially expressed donor region genes with whole genome microarrays using RNA from eight obesity tissues (Aim 3), sequence selected donor region genes (Aim 4), determine cis or trans control of mRNA levels for differentially expressed genes (Aim 5), produce transgenic mice overexpressing ten candidate genes prioritized by the results of Aims 3-5 (Aim 6) and identify microRNAs or other transcribed non-coding genes that may influence obesity (Aim 7). The long term outcome is that we will identify at least one novel and strong and plausible obesity gene.

描述（由申请人提供）：定位克隆-通过将映射的肥胖性状和基因重合到特定染色体区域来鉴定肥胖基因，提供了发现新的肥胖基因和对引起肥胖的机制进行新的了解的有力方法。小鼠已被用于首次识别比任何其他模型更多的人类肥胖基因。本研究的目的是利用定位克隆的方法来鉴定小鼠同源品系中的肥胖基因。同源小鼠品系与背景品系相同，除了来自供体品系的染色体区域。同源和背景之间的表型差异是由于供体菌株的等位基因与背景菌株的功能效应不同。对该提案做出贡献的两个实验室已经在小鼠2号染色体上与人类20号染色体同源的区域中鉴定出了几种具有统计学显着肥胖表型的小鼠同类品系，其中已经绘制了几个肥胖数量性状基因座。我们的一般假设是，在同源供体区域的一个或多个基因或转录影响肥胖。我们具体的工作假设是，肥胖症的基因/转录本可以确定为大多数同类。我们提出了七个具体目标。我们将：确定饮食、性别和年龄对肥胖的影响（目标1），通过用保留肥胖表型的更小供体区域培育同源物来鉴定含有肥胖基因的最小染色体基因座（目标2），使用来自八种肥胖组织的RNA用全基因组微阵列发现差异表达的供体区域基因（目标3），对选择的供体区域基因进行测序（目标4），确定差异表达基因的mRNA水平的顺式或反式控制（目标5），产生过表达根据目标3-5的结果优先的10个候选基因的转基因小鼠（目标6），并鉴定可能影响肥胖的microRNA或其他转录的非编码基因（目标7）。长期的结果是，我们将确定至少一个新的和强大的和合理的肥胖基因。