利用体细胞镶嵌变异回溯人类大脑中的细胞系统发育
Backtracking Cell Phylogenies in the Human Brain with Somatic Mosaic Variants.
作者信息
Bizzotto Sara
机构信息
Sorbonne Université, Institut du Cerveau (Paris Brain Institute) ICM, Inserm, CNRS, Hôpital de la Pitié Salpêtrière, Paris, France.
Université Paris Cité, Imagine Institute, Paris, France.
出版信息
Methods Mol Biol. 2025;2886:201-220. doi: 10.1007/978-1-0716-4310-5_10.
Somatic mosaic variants, and especially somatic single nucleotide variants (sSNVs), occur in progenitor cells in the developing human brain frequently enough to provide permanent, unique, and cumulative markers of cell divisions and clones. Here, we describe an experimental workflow to perform lineage studies in the human brain using somatic variants. The workflow consists in two major steps: (1) sSNV calling through whole-genome sequencing (WGS) of bulk (non-single-cell) DNA extracted from human fresh-frozen tissue biopsies, and (2) sSNV validation and cell phylogeny deciphering through single nuclei whole-genome amplification (WGA) followed by targeted sequencing of sSNV loci.
体细胞镶嵌变异，尤其是体细胞单核苷酸变异（sSNV），在发育中的人类大脑的祖细胞中频繁出现，足以提供细胞分裂和克隆的永久、独特和累积标记。在这里，我们描述了一种使用体细胞变异在人类大脑中进行谱系研究的实验工作流程。该工作流程包括两个主要步骤：（1）通过对从人类新鲜冷冻组织活检中提取的大量（非单细胞）DNA进行全基因组测序（WGS）来调用sSNV，以及（2）通过单核全基因组扩增（WGA），然后对sSNV位点进行靶向测序来验证sSNV并解读细胞系统发育。
Zotero 插件