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

靶向重组以查明小鼠大易感位点内的负责区域

• 批准号: 10021676
• 负责人: Galina Petukhova
• 金额: $ 15.25万
• 依托单位: HENRY M. JACKSON FDN FOR THE ADV MIL/MED
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-20 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10021676
• 关键词: Animal Model; Back; Bacterial Artificial Chromosomes; Base Pairing; Biochemical; DNA Double Strand Break; DNA Repair Pathway; Development; Disease; Elements; Future; Gametogenesis; Generations; Genes; Genetic Polymorphism; Genetic Recombination; Guide RNA; Health; Human; Location; Meiosis; Mouse Strains; Mus; Nonhomologous DNA End Joining; Pattern; Phenotype; Predisposition; Recombinants; Route; Site; Sorting - Cell Movement; Susceptibility Gene; Therapeutic Intervention; Time; Untranslated RNA; Work; base; causal variant; cost efficient; disease phenotype; functional genomics; gene product; genome wide association study; homologous recombination; knockout animal; novel strategies; novel therapeutics; positional cloning; reference genome; repaired; therapeutic development; tool; trait; vector

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.

抽象的 位置克隆以识别负责疾病或特定性状的基因涉及 通过人类的全基因组关联研究和 重组菌株和小鼠背面的表型分析。然后负责的基因 在此基因座中，根据其表达模式，生化活动的确定 相应基因敲除动物模型的基因产物和表型。很遗憾， 大尺寸的敏感性基因座（通常> 5 MB）使下游调查步骤 相当具有挑战性。到目前为止，缩小负责任地区范围的唯一方法是退缩 - 越过载体小鼠，以通过同源重组来修剪基因座。但是， 自发重组的机会很低，将基因座缩小到可管理的尺寸 通常不可行。我们建议开发人为诱导重组的方法 在易感基因座（靶向重组）内的定义位置。这将提供 可靠的途径，以查明病因突变，最终是该疾病的机制 有问题。