Targeted recombination to pinpoint responsible regions within large susceptibility loci in mice

摘要 通过定位克隆来确定疾病或特定性状的基因， 通过人类全基因组关联研究绘制易感基因位点， 重组菌株的表型分析和小鼠中的回交。那么负责的基因 基于它们的表达模式、这些基因座内的生物化学活性， 相应敲除动物模型的基因产物和表型。不幸的是， 易感基因座的大尺寸（通常>5 Mb）使得下游研究步骤 相当具有挑战性。到目前为止，缩小责任区域的唯一方法是支持- 将携带者小鼠杂交，以便通过同源重组来修剪基因座。但 自发重组的机会非常低，将基因座修剪到可管理的大小 通常不可行。我们建议开发人工诱导重组的方法 在易感性基因座内的确定位置（靶向重组）。这将提供 一种可靠的途径来确定致病突变，并最终确定疾病的机制。 有问题。

ABSTRACT Positional cloning to identify the genes responsible for disease or particular traits involves mapping of the susceptibility locus through genome-wide association studies in humans and phenotypic analysis of recombinant strains and back-crossing in mice. Then responsible genes within such loci are identified based on their expression patterns, biochemical activities of the gene products and phenotypes of the corresponding knockout animal models. Unfortunately, the large size of susceptibility loci (usually >5 Mb) makes the downstream investigative steps rather challenging. The only way to narrow down the responsible region so far was to back- cross the carrier mice in order to trim the locus by homologous recombination. However, the chance of spontaneous recombination is very low, and trimming the locus to a manageable size is usually not feasible. We propose to develop the approach to artificially induce recombination at defined locations within the susceptibility locus (targeted recombination). This will provide the reliable route to pinpoint the causative mutations and ultimately, the mechanism of the disease in question.