Engineering of complex alleles

与人类疾病相关的突变可以被改造成小鼠的同等基因，由此产生的转基因小鼠为疾病机制提供了重要的见解，为未来的药物提供了新的靶点。此外，这些人类疾病的小鼠模型在新药的临床前测试中具有相当大的实用价值，并可以帮助开发新的工具来帮助疾病诊断。到目前为止，使用小鼠模型探索的大多数疾病突变仅限于影响编码蛋白质的DNA的突变。然而，过去十年对人类遗传变异的研究表明，非蛋白质编码DNA序列的突变和变异以及人类基因的整体结构在定义疾病和疾病风险方面发挥着重要作用。例如，基因的三维结构和它在一个更大的结构域(从10‘S到100’S的几千个DNA序列碱基)中的位置对基因调控产生了巨大的影响。这可能会被该结构域结构的巨大变化或干扰该结构域边界的较小突变所干扰。我们需要开发工具，能够在比常规操作更大的规模上操纵和评估DNA序列，以便在小鼠身上对人类疾病进行准确的调查和建模。小鼠和人类在这些非编码序列和整体基因结构上也存在显著差异。因此，需要对小鼠的人类疾病突变进行更复杂的建模。这对于在老鼠身上准确地模拟人类疾病，以及测试作用于人类基因产品的新药或甚至对人类基因序列起作用的新一代疗法都是重要的。世界各地的一些研究团队已经报告了这些方法，但还没有明确的最佳实践来以这种方式设计小鼠基因组。英国研究人员能够通过其大学内部的核心设施或通过获得国家和国际计划来获得实现简单基因组修改的技术。然而，英国目前没有能力在老鼠身上进行更复杂的大规模工程，这是我们对疾病生物学的理解现在所需要的。我们的集群将解决这一未得到满足的需求。我们将探索和优化不同的实验参数，并比较不同的方法。我们的目标是建立强大的管道，将完整的人类基因和染色体片段工程到小鼠体内。我们将建立概念验证模型，包括这项技术的关键应用，实现更好的疾病模型，并帮助我们理解癌症、血液学和神经退化领域的疾病生物学。这项技术和由此产生的模型将有助于我们对疾病的理解和新疗法的开发。

Mutations associated with human disease can be engineered into the equivalent genes in mice, and the resulting genetically modified mice provide important insights into disease mechanism, revealing new targets for future medicines. Furthermore, these mouse models of human disease have considerable utility for preclinical testing of new medicines and can help the development of new tools to help disease diagnosis. To date, most disease mutations that have been explored using mouse models are restricted to mutations which affect the DNA encoding proteins. Investigations into genetic variation in humans over the last decade, however, have revealed that mutations and variation in non-protein-coding DNA sequences and the overall structure of human genes play an important role in defining disease and disease-risk. For example, the three dimensional architecture of a gene and its location within a larger domain (spanning 10's to 100's of thousands of DNA sequence bases) impacts gene regulation enormously. This can be perturbed by large variations in the structure of this domain or smaller mutations that interfere with the domain's boundaries. We need to develop tools that enable manipulation and assessment of DNA sequences at much larger scale then conventionally performed to allow an accurate investigation and modelling of human disease in mouse. Significant differences also exist between mouse and human in these non-coding sequences and overall gene structure. Consequently, more sophisticated modelling of human disease mutations in the mouse is required. This is important both with respect to accurately modelling the human disease in mouse but also for testing new medicines that act on human gene products or new generation therapies that even on human gene sequences.Methods have been reported by a few research teams around the world, but there is no clear best practice established for engineering the mouse genome in this way. UK researchers are able to access technologies for achieving simple modifications of the genome, through core facilities within their universities or via access to national and international programmes. There is, however, currently no capacity within the UK for the more sophisticated large-scale engineering in the mouse that our understanding of disease biology now demands.Our cluster will address this unmet need. We will explore and optimize the different experimental parameters and compare different methodologies. We aim to establish robust pipelines for engineering complete human genes and chromosomal segments into the mouse. We will establish proof-of-concept models which encompass key applications of this technology, achieving better disease models and helping our understanding of the biology of disease in areas of cancer, haematology and neurodegeneration. The technology and the resulting models will assist in our understanding of disease and the development of new therapies.