通过具有谱系限制驱动因子的双斑MARCM在单细胞分辨率下进行谱系分析。

Lineage Analysis at Single-Cell Resolution by Twin-Spot MARCM with Lineage-Restricted Drivers.

作者信息

Previero Angelica, Yang Ching-Po, Lee Tzumin

机构信息

Life Science Institute, University of Michigan, Ann Arbor, MI, USA.

Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, MI, USA.

出版信息

Methods Mol Biol. 2025;2886:401-420. doi: 10.1007/978-1-0716-4310-5_20.

DOI:10.1007/978-1-0716-4310-5_20

PMID:39745652

Abstract

Cell lineage analysis is primarily undertaken to understand cell fate specification and diversification along a cell lineage tree. Built with dual repressible markers, twin-spot mosaic analysis with repressible cell markers (MARCM) labels the two daughter cells made by a common precursor in distinct colors. The power of twin-spot MARCM to systematically subdivide complex lineages is exemplified in studies of Drosophila neural stem-cell lineages. To achieve targeted lineage analysis, lineage-restricted drivers can be created by restricting and immortalizing gene expression to a lineage of interest. Here, we describe detailed methods to perform lineage analysis in Drosophila neural stem-cell lineages utilizing lineage-specific drivers. Our innovative lineage tracing method can help resolve complex tissue development.

摘要

细胞谱系分析主要用于了解细胞命运特化以及沿细胞谱系树的多样化过程。利用双可抑制标记构建的、带有可抑制细胞标记的双斑镶嵌分析（MARCM），会以不同颜色标记由共同前体细胞产生的两个子细胞。双斑MARCM在系统细分复杂谱系方面的能力在果蝇神经干细胞谱系研究中得到了体现。为了实现靶向谱系分析，可以通过将基因表达限制并永生化到感兴趣的谱系中来创建谱系限制驱动子。在此，我们描述了利用谱系特异性驱动子在果蝇神经干细胞谱系中进行谱系分析的详细方法。我们创新的谱系追踪方法有助于解析复杂的组织发育过程。