NIH Director's Pioneer Award

翻译后摘要：细胞命运图描述了细胞分裂，迁移和凋亡的顺序如何改变受精卵 是理解干细胞生物学的基础。然而，它只存在于透明蠕虫C中。 在这种情况下，对每一次细胞分裂的繁琐的显微镜观察允许构建一个完整的 细胞命运图更复杂和不透明动物证明是不太顺从的。然而DNA复制不可避免地 产生体细胞突变。因此，多细胞生物体包括马赛克，其中大多数细胞 获得独特的基因组，有可能描绘他们的祖先。我们建议建造 哺乳动物细胞命运图使用系统发育方法被动追溯胚胎关系， 从而推断出在发育过程中发生突变的顺序。我们发现聚鸟嘌呤 重复的DNA序列是特别有用的遗传标记，因为它们经常在过程中改变长度。 分裂。为了证明其可行性，我们利用遗传学方法重建了培养小鼠的谱系 NIH3T3成纤维细胞基于影响聚鸟嘌呤标记物长度的突变。然后我们雇用了 全基因组扩增以在取自小鼠的单细胞中对聚鸟嘌呤标记进行基因分型， 胚胎遗传学来推断取样组织的发育关系。我们的初步结果 证明了这种方法用于回顾性地产生完整的哺乳动物细胞命运的潜力， 原则上，它可以描述任何细胞的发育谱系，并解决与以下相关的悬而未决的问题： 干细胞生物学

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.