TRANSMITOCHONDRIAL MOUSE MODELS OF HUMAN DISEASE

描述(由申请人提供)：该项目旨在开发一种 人类线粒体疾病的新小鼠模型。基因工程和 将利用分子生物学技术创造异质性 携带突变线粒体基因组的“传递线粒体”小鼠。这款车 而线粒体转移的过程将是相当重要的 对我们理解特定的线粒体突变的重要性，如 并导致人类新策略和治疗方法的发展 受线粒体功能或功能异常影响的代谢性疾病 突变。在试点实验中，创造传递线粒体的能力 将异质状态传递给子代的小鼠模型 血统被鉴定出来。随着基因转移技术和 基于聚合酶链式反应的程序，这个项目将以技术开发为目标 建立定向线粒体DNA(MtDNA)小鼠模型 删除。具体目标包括：(1)发展和优化 线粒体转染法，(2)转染型线粒体 (或细胞)进入小鼠卵子以产生异质传递线粒体小鼠 这将概括与人类线粒体DNA疾病相关的缺失- 没有可供研究的动物模型，以及(3)表征 在这个项目的过程中创建的线粒体小鼠血统。 异质传递线粒体缺失突变体的产生将是一个 关键的第一步，促进线粒体功能和 疾病的发展。最初，这个模型将用于探索疾病 体内系统中的发病机制和线粒体动力学。最终， 小鼠线粒体传递模型将被用来探索 线粒体基因组在人类代谢性疾病过程中的作用 开发新的人类基因疗法。

DESCRIPTION (Provided by applicant): This project is designed to develop a novel mouse model of human mitochondrial disease. Genetic engineering and molecular biological techniques will be utilized to create heteroplasmic "transmitochondrial" mice harboring a mutant mitochondrial genome. This model and the procedures for mitochondrial transfer will be of considerable importance toward our understanding of a specific mitochondrial mutation, as well as leading to the development of novel strategies and therapies for human metabolic diseases influenced by aberrations in mitochondrial function or mutation. In pilot experiments, the ability to create transmitochondrial mouse models that transmit the heteroplasmic state to offspring in maternal lineages was identified. With the advent of gene transfer technologies and PCR-based procedures, this project will target the development of technology to establish a mouse model harboring a directed mitochondria DNA (mtDNA) deletion. Specific Aims include: (1) development and optimization of mitochondria transfection procedures, (2) transfer of transfected mitochondria (or cells) into mouse ova to produce heteroplasmic transmitochondrial mice that will recapitulate a deletion associated with human mtDNA-based disease - for which no animal model exists for study, and (3) characterization of transmitochondrial mouse lineages created over the course of this project. Production of heteroplasmic transmitochondrial deletion mutants will be a critical first step, facilitating the study of mitochondrial function and disease progression. Initially, this model will serve to explore disease pathogenesis and mitochondrial dynamics in an in vivo system. Ultimately, transmitochondrial mouse models will be used to explore the role of the mitochondrial genome in human metabolic disease processes and in the development of novel human gene therapies.