TRANSMITOCHONDRIAL MOUSE MODELS OF HUMAN DISEASE

描述（由申请人提供）：本项目旨在开发一个 人类线粒体疾病的新型小鼠模型。 遗传工程和 分子生物学技术将被用来创造异质 携带突变线粒体基因组的“线粒体传递”小鼠。 该模型 线粒体转移的过程将是相当大的 对我们理解特定线粒体突变的重要性， 并导致开发新的策略和疗法， 受线粒体功能异常影响的代谢性疾病，或 突变 在初步实验中，创造线粒体传递的能力 将异质状态传递给子代的小鼠模型， 血统被确认。 随着基因转移技术的出现， 基于PCR的程序，本项目将针对技术开发 建立一种携带定向线粒体DNA（mtDNA）的小鼠模型， 删除。 具体目标包括：（1）开发和优化 线粒体转染程序，（2）转染的线粒体的转移 (or细胞）进入小鼠卵中以产生异质性线粒体小鼠 它将重现与人类mtDNA疾病相关的缺失- 没有动物模型可供研究，和（3）表征 在这个项目的过程中创建的线粒体小鼠谱系。 异质性线粒体缺失突变体的产生将是一个重要的研究方向。 关键的第一步，促进线粒体功能的研究， 疾病进展。 最初，该模型将用于探索疾病 在体内系统中的发病机制和线粒体动力学。 最后， transmitochondrial小鼠模型将被用来探索的作用， 线粒体基因组在人类代谢疾病过程中的作用， 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.