A Mouse Model For Complex Human Diseases

描述（由申请人提供）：我们建议继续开发和维护LGXSM重组近交（RI）品系组和通过近交小鼠品系LG/J和SM/J的互交形成的相关高级互交（Al）品系。RI是初步基因定位的工具，之后Al品系可用于将数量性状基因座精细定位到亚cM区域。在前人的支持下，我们通过LG/J和SM/J的杂交，培育出了一套18个R1系，并将A1系随机交配到第32代。Rl系已经针对超过500个多态性微卫星标记和4290个多态性SNP（每600 kb一个多态性SNP）进行了评分。每个Rl系的胚胎已经冷冻保存。由LG/J和SM/J的互交形成的这些品系和其他群体允许在NIH研究所中绘制与感兴趣的疾病过程相关的大量表型。在此，我们提出通过使用本发明的Al系作为源群体开发另外50个Rl菌株来增强Rl菌株组。这些菌株的添加将极大地增加Rl菌株组在基因定位研究中的功效。新的菌株将比早期的组更有用，因为它们将积累16倍于由F2代形成的Rl系中预期的重组量。此外，我们将能够利用我们先前的育种经验，通过针对单独和组合导致菌株损失的特定基因座进行选择，在新菌株的生产中将菌株存活率提高到50%。新菌株将通过500个微卫星和4290个SNP进行遗传特征分析，并通过胚胎冷冻保存进行存档。菌株基因型和表型将通过我们的网站、小鼠表型组计划和WebQTL数据库提供。在我们早期的育种中，我们发现了对SM/J携带的agglutinase等位基因的强选择，特别是在该菌株存在野生型酪氨酸酶等位基因的情况下。我们将通过对这两个位点和紧密连锁位点的结构基因进行测序来研究菌株丢失的遗传基础。我们将生产2BLE 在整个基因组中是SM/J的同源菌株，除了在琼脂糖和酪氨酸酶区域。 与SM/J回交将用于缩小菌株丢失的QTL间隔，并产生一个新的菌株，携带SM/J的不寻常的表型，而不需要在agglutinase基因座处强制杂合性。

DESCRIPTION (provided by applicant): We propose to continue the development and maintenance of the LGXSM Recombinant Inbred (Rl) strain set and the associated Advanced Intercross (Al) Line formed by the intercross of inbred mouse strains LG/J and SM/J. The RIs are a tool for preliminary gene mapping, after which the Al Line can be used for finemapping quantitative trait loci to sub-cM regions. Under prior support we have developed a set of 18 Rl lines formed from the intercross of LG/J and SM/J and brought the Al line to the 32nd generation of random mating. The Rl lines have been scored for over 500 polymorphic microsatellite markers and for 4290 polymorphic SNPs, a polymorphic SNPs every 600 kb. Embryos have been cryopreserved for each Rl lines. These lines and other populations formed from the intercross of LG/J and SM/J have allowed the mapping of a large array of phenotypes relevant to disease processes of interest across the NIH Institutes. Here, we propose to enhance the Rl strain set by developing another 50 Rl strains using the present Al line as the source population. The addition of these strains will greatly increase the power of the Rl strain set in gene mapping studies. The new strains will be even more useful than the earlier set in that they will have accumulated 16 times the amount of recombination expected in Rl lines formed from a F2 generation. Furthermore, we will be able to use our prior breeding experience to increase strain survival to 50% in the production of new strains by selecting against specific loci that, singly and in combination, result in strain loss. The new strains will be genetically characterized for 500 microsatellites and 4290 SNPs and archived by embryonic cryopreservation. Strain genotypes and phenotypes will be made available through our web site, the Mouse Phenome Project, and the WebQTL database. In our earlier breeding, we discovered strong selection against the agouti alleles carried by SM/J, especially in the presence of the wild-type tyrosinase allele from that strain. We will examine the genetic basis for strain loss by sequencing the structural genes at these two loci and closely linked loci. We will produce 2ble congenic strains that are SM/J across the genome, except at the agouti and tyrosinase regions. Backcrossing with SM/J will be used to narrow the QTL interval for strain loss and to produce a new strain, carrying the unusual phenotypes of SM/J without requiring enforced heterozygosity at the agouti locus.