Department of Cell Biology, Emory University School of Medicine, Atlanta, GA, USA.
Methods Mol Biol. 2023;2677:61-79. doi: 10.1007/978-1-0716-3259-8_3.
The last several years have seen an increasing number of examples of transgenerational epigenetic inheritance, in which phenotypes are inherited for three or more generations without changes to the underlying DNA sequence. One model system that has been particularly useful for studying transgenerational epigenetic inheritance is C. elegans. Their short generation time and hermaphroditic reproduction have allowed multiple transgenerational phenotypes to be identified, including aging, fertility, and behavior. However, it is still not clear how transgenerational epigenetic inheritance from the germline affects embryogenesis. Fortunately, the C. elegans embryo has a unique property that makes it ideal for addressing this question: they develop via an invariant lineage, with each cell undergoing stereotypical cell divisions to adopt the same cell fate in every individual embryo. Because of this invariant cell lineage, automated lineage tracing and single-cell RNA-seq can be employed to determine how transgenerational epigenetic inheritance from the germline affects developmental timing and cell fate. Unfortunately, difficulties with these techniques have severely limited their adoption in the community. Here, we provide a practical guide to automated lineage tracing coupled with single-cell RNA-seq to facilitate their use in studying transgenerational epigenetic inheritance in C. elegans embryos.
在过去的几年中，越来越多的跨代表观遗传遗传实例被发现，其中表型在不改变底层 DNA 序列的情况下被遗传了三代或更多代。在研究跨代表观遗传遗传方面，一个特别有用的模型系统是秀丽隐杆线虫。它们较短的世代时间和雌雄同体的繁殖方式允许鉴定出多种跨代表型，包括衰老、生育能力和行为。然而，目前仍不清楚来自生殖系的跨代表观遗传遗传如何影响胚胎发生。幸运的是，秀丽隐杆线虫的胚胎具有一个独特的特性，使其成为解决这个问题的理想选择：它们通过不变的谱系发育，每个细胞都经历典型的细胞分裂，从而在每个个体胚胎中采用相同的细胞命运。由于这种不变的细胞谱系，可以采用自动谱系追踪和单细胞 RNA-seq 来确定来自生殖系的跨代表观遗传遗传如何影响发育时间和细胞命运。不幸的是，这些技术的困难严重限制了它们在该领域的应用。在这里，我们提供了一个实用的指南，介绍如何将自动谱系追踪与单细胞 RNA-seq 相结合，以促进它们在研究秀丽隐杆线虫胚胎中跨代表观遗传遗传的应用。
