用体细胞镶嵌变异回溯人脑细胞系统发育。

Q4 Biochemistry, Genetics and Molecular Biology

Methods in molecular biology Pub Date : 2025-01-01 DOI:10.1007/978-1-0716-4310-5_10

Sara Bizzotto

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引用次数: 0

摘要

体细胞镶嵌变异，尤其是体细胞单核苷酸变异（sSNVs），经常发生在发育中的人脑祖细胞中，足以为细胞分裂和克隆提供永久的、独特的和累积的标记。在这里，我们描述了一个实验工作流程，在人类大脑中使用体细胞变异进行谱系研究。该工作流程包括两个主要步骤：(1)通过从人类冷冻组织活检中提取的大量（非单细胞）DNA的全基因组测序（WGS）来调用sSNV；(2)通过单核全基因组扩增（WGA）来验证sSNV并进行细胞系统发育解码，然后对sSNV位点进行靶向测序。

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Backtracking Cell Phylogenies in the Human Brain with Somatic Mosaic Variants.

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.

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来源期刊

Methods in molecular biology Biochemistry, Genetics and Molecular Biology-Genetics

CiteScore

2.00

自引率

0.00%

发文量

3536

期刊介绍： For over 20 years, biological scientists have come to rely on the research protocols and methodologies in the critically acclaimed Methods in Molecular Biology series. The series was the first to introduce the step-by-step protocols approach that has become the standard in all biomedical protocol publishing. Each protocol is provided in readily-reproducible step-by-step fashion, opening with an introductory overview, a list of the materials and reagents needed to complete the experiment, and followed by a detailed procedure that is supported with a helpful notes section offering tips and tricks of the trade as well as troubleshooting advice.