NIH Director's Pioneer Award

NIH 院长先锋奖

• 批准号: 7919259
• 负责人: MARSHALL S. HORWITZ
• 金额: $ 78万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-30 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7919259
• 关键词: Adult; Affect; Apoptosis; Award; Cell division; Cells; DNA Sequence; DNA biosynthesis; Development; Embryo; Fibroblasts; Genetic Markers; Genome; Genotype; Length; Mammalian Cell; Maps; Microscopic; Mitosis; Mus; Mutation; Organism; Phylogenetic Analysis; Somatic Mutation; Tissue Sample; United States National Institutes of Health; abstracting; base; migration; stem cell biology; zygote; Adult; Affect; Apoptosis; Award; Cell division; Cells; DNA Sequence; DNA biosynthesis; Development; Embryo; Fibroblasts; Genetic Markers; Genome; Genotype; Length; Mammalian Cell; Maps; Microscopic; Mitosis; Mus; Mutation; Organism; Phylogenetic Analysis; Somatic Mutation; Tissue Sample; United States National Institutes of Health; abstracting; base; migration; stem cell biology; zygote

Abstract: Cell fate maps describe how the sequence of cell division, migration, and apoptosis transform a zygote into an adult and are fundamental to understanding stem cell biology. Yet, it is only in the transparent worm C. elegans where tedious microscopic observation of each cell division has allowed for construction of a complete cell fate map. More complexand opaqueanimals prove less yielding. DNA replication, however, inevitably generates somatic mutations. Consequently, multicellular organisms comprise mosaics where most cells acquire unique genomes that are potentially capable of delineating their ancestry. We propose to construct mammalian cell fate maps using a phylogenetic approach to passively retrace embryonic relationships by deducing the order in which mutations have arisen during development. We have found that polyguanine repeat DNA sequences are particularly useful genetic markers, because they frequently change length during mitosis. To demonstrate feasibility, we have used phylogenetics to reconstruct the lineage of cultured mouse NIH3T3 fibroblasts based on mutations affecting the length of polyguanine markers. We have then employed whole genome amplification to genotype polyguanine markers in single cells taken from a mouse and used phylogenetics to infer the developmental relationships of the sampled tissues. Our preliminary results demonstrate the potential of this approach for retrospectively producing a complete mammalian cell fate that, in principle, could describe the developmental lineage of any cell and resolve outstanding questions relevant to stem cell biology.

摘要：细胞命运图描述了细胞分裂，迁移和凋亡的序列如何改变合子 成人，是理解干细胞生物学的基础。但是，只有在透明的蠕虫中。 秀丽隐杆线虫对每个细胞分裂的乏味的微观观察都可以构建完整的 细胞命运图。更复杂，不透明的动物证明产量较低。但是，DNA复制不可避免地 产生体细胞突变。因此，多大多数细胞的多细胞生物包括镶嵌物 获取潜在能够描绘其血统的独特基因组。我们建议建造 使用系统发育方法的哺乳动物细胞命运图，通过 推断出在发育过程中突变产生的顺序。我们发现多瓜氨酸 重复DNA序列是特别有用的遗传标记，因为它们在 有丝分裂。为了证明可行性，我们已经使用系统发育学来重建培养的小鼠的谱系 NIH3T3成纤维细胞基于影响多圭胺标记长度的突变。然后我们就雇用了 从小鼠采集的单个细胞中，整个基因组扩增到基因型的多圭亚氨酸标记物并使用 系统发育学来推断采样组织的发育关系。我们的初步结果 证明这种方法回顾性产生完整的哺乳动物细胞命运的潜力，该命运 原则上，可以描述任何细胞的发展谱系，并解决与 干细胞生物学。