Mendelian inheritance of artificial chromosomes

人工染色体的孟德尔遗传

• 批准号: 10666591
• 负责人: Ben E. Black
• 金额: $ 116.96万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-10 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10666591
• 关键词: Address; Animal Model; Animals; Area; Artificial Chromosomes; Artificial Mammalian Chromosomes; Behavior; Biological Models; Biotechnology; Cell division; Centromere; Chromatin; Chromosomal Stability; Chromosome Pairing; Chromosomes; Circular DNA; Clinic; Custom; DNA; DNA Sequence; Data; Development; Elements; Embryo; Ensure; Epigenetic Process; Eukaryota; Female; Foundations; Genes; Genetic; Genetic Recombination; Genome; Genome Components; Genome engineering; Germ Cells; Goals; Harvest; Histones; Human Chromosomes; Human Genome Project; In Vitro; Industrialization; Inherited; Investments; Laws; Mammalian Cell; Mammalian Chromosomes; Mammals; Medicine; Meiosis; Mitosis; Mitotic; Mitotic Cell Cycle; Mus; Nucleic Acid Sequence Homology; Nucleosomes; Organ Transplantation; Outcome; Patients; Privatization; Prophase; Research Personnel; Research Project Grants; Saccharomycetales; Sex Chromosomes; Sexual Reproduction; Source; Specific qualifier value; Technology; Testing; Time; Variant; Work; Writing; Yeasts; centromere protein A; design; drug development; frontier; genetic element; genetic payload; in vivo; innovation; insight; mouse model; quantum; segregation; success; synthetic biology; synthetic construct; telomere; tool; transmission process; vector

Synthetic mammalian artificial chromosomes (MACs) represent a new frontier in genome technology, with the potential to transform chromosome and synthetic biology and stimulate the development of numerous radical advances in medicine. Human Genome Project-Write aims to generate an entire set of synthetic human chromosomes. Short of this ambitious goal, MACs have enormous potential for breakthroughs in biotechnology and medicine, such as creating humanized animal models for drug development or for harvesting patient- personalized organs for transplantation. Furthermore, building MACs from minimal components will advance our fundamental understanding of what comprises a mammalian chromosome. As vehicles for genetic inheritance, fully functional chromosomes are faithfully transmitted through mitosis and the specialized meiotic divisions underlying eukaryotic sexual reproduction and Mendelian inheritance. Our goal is to construct the first MACs that achieve faithful inheritance through the germline, using mouse as a model system. One obstacle is the centromere, the locus on each chromosome that directs transmission through both mitosis and meiosis. Because mammalian centromeres are not encoded in the DNA sequence, it is unclear how to build synthetic chromosomes containing this crucial element. There are additional challenges to create MACs that pair and recombine as homologous chromosomes in meiosis. To solve these problems, we will hijack the existing cellular machinery for assembling centromere chromatin and incorporate additional genetic elements to ensure meiotic pairing and recombination. This effort requires innovation at multiple levels: designing MAC vectors encoding key functional elements, assembling large synthetic DNA constructs, and ultimately creating animals to test MACs in vivo. The proposed work builds on recent advances from the co-investigators’ teams in all of these areas, and we have key tools and expertise in place to build the necessary DNA templates, introduce them into embryos, analyze the outcomes, and develop alternative strategies as necessary. The most meaningful preliminary data would be to show a synthetic artificial chromosome that is successfully transmitted through mitosis and meiosis in vivo, but achieving this step is a major goal of our proposal and will require substantial investment of time and effort. Thus, we are requesting support for this project without the preliminary data that would demonstrate high likelihood of success, justifying consideration of our proposal as part of the T-R01 mechanism. We use mouse as a relatively rapid and tractable mammalian model system with outstanding opportunities for testing and debugging MACs, and our advances should readily transfer to other species for applications in biotechnology and medicine. Success in this project will represent a quantum leap in the development of synthetic artificial chromosome that are fully functional in vivo, providing unprecedented genome engineering capabilities in animal models and enabling diverse synthetic biology applications.

合成哺乳动物人工染色体（MAC）代表基因组技术的新边界， 有可能改变染色体和合成生物学并刺激众多的发展 激进的医学进步。人类基因组项目 - 旨在产生一组合成人类 染色体。除了这个雄心勃勃的目标之外，MAC具有巨大的生物技术突破潜力 和医学，例如为药物开发或收集患者创建人性化动物模型 - 个性化器官进行移植。此外，从最小组件中构建Mac将推进 我们对包含哺乳动物染色体的内容的基本理解。 作为遗传遗传的车辆，通过 有丝分裂和真核有性繁殖的专门减数分裂分裂和Mendelian 遗产。我们的目标是构建通过种系实现忠实继承的第一批Mac，使用 鼠标作为模型系统。一个障碍是丝粒，每个染色体的座位 通过有丝分裂和减数分裂的传播。因为哺乳动物的丝粒未编码在DNA中 序列，尚不清楚如何构建包含此关键元素的合成染色体。有 在减数分裂中，创建将MAC与同源染色体配对并重组的MAC面临的其他挑战。到 解决这些问题，我们将劫持现有的蜂窝机械来组装中心粒染色质和 纳入其他遗传元素，以确保减数分裂和重组。 这项工作需要在多个层面上进行创新：设计编码密钥功能的Mac向量 元素，组装大型合成DNA构建体，并最终创建动物在体内测试MAC。 拟议的工作是建立在所有这些领域的共同投资者团队的最新进展的基础上的，我们 拥有关键工具和专业知识来构建必要的DNA模板，将其引入胚胎， 分析结果，并根据需要制定替代策略。最有意义的初步数据 将显示一个合成的人造染色体，该染色体通过有丝分裂和减数分裂成功地传播 体内，但是实现这一步骤是我们提议的主要目标，将需要大量时间投资 和努力。这是我们要求对该项目的支持，而没有初步数据 成功的可能性很大，证明我们的提议是T-R01机制的一部分。 我们将鼠标用作相对较快且可进行的哺乳动物模型系统，并具有出色的 测试和调试Mac的机会，我们的进步应轻易转移到其他物种 生物技术和医学中的应用。该项目的成功将代表 开发体内功能齐全的合成人造染色体，提供前所未有的 动物模型中的基因组工程能力并实现潜水员合成生物学应用。