A multi-kingdom genetic barcoding system for precise clone isolation

doi:10.1038/s41587-025-02649-1

2025-05-21

Anne‐Sophie Archambault

Ramon I. Klein Geltink

DOI：10.1038/s41587-025-02649-1

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摘要

En 中文

基于DNA条形码的细胞标记策略已能够分析异质群体中的克隆大小动态及克隆特异性的转录组景观。然而，从复杂群体中分离表现出特定表型的目标克隆仍然具有挑战性。在此，我们介绍一种多界域遗传条形码系统CloneSelect，该系统可通过条形码特异性的CRISPR碱基编辑触发目标细胞克隆表达报告基因以实现分离。在CloneSelect中，细胞首先被稳定标记上DNA条形码并扩增，使其亚群可接受给定实验。在特定时间表现出感兴趣表型或基因型的克隆，随后可通过CRISPR碱基编辑从实验期间储存的初始或后续细胞池中分离出来。CloneSelect具有可扩展性，且与单细胞RNA测序兼容。我们通过人胚肾293T细胞、小鼠胚胎干细胞、人 pluripotent stem cells、酵母细胞及细菌细胞展示了CloneSelect的通用性。

Keyword:

EPIGENETIC MEMORY

LINEAGE

CELLS

DIFFERENTIATION

DNA

ADAPTATION

INDUCTION

DYNAMICS

DESIGN

FATE

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Nature Biotechnology

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41.7

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