英属哥伦比亚大学Nozomu Yachie小组取得一项新突破。他们报道了用于精确克隆分离的多界遗传条形码系统。相关论文于2025年5月21日发表于国际顶尖学术期刊《自然—生物技术》杂志上。

在这里，该课题组提出了一个多王国遗传条形码系统，CloneSelect，它能够通过条形码特异性CRISPR碱基编辑触发靶细胞克隆表达报告基因进行分离。在CloneSelect中，细胞首先用DNA条形码稳定地标记，然后进行繁殖，这样它们的亚群就可以接受给定的实验。然后，可以从CRISPR碱基编辑实验期间存储的初始或后续细胞池中分离出在给定时间显示感兴趣的表型或基因型的克隆。CloneSelect是可扩展的，与单细胞RNA测序兼容。小组证明了CloneSelect在人胚胎肾293T细胞、多主题胚胎干细胞、人多能干细胞、酵母细胞和细菌细胞中的多功能性。

据介绍，利用DNA条形码的细胞标记策略可以分析异种群体的克隆大小动态和克隆限制性转录组景观。然而，从复杂的种群中分离出具有特定表型的目标克隆仍然具有挑战性。

附：英文原文

Title: A multi-kingdom genetic barcoding system for precise clone isolation

Author: Ishiguro, Soh, Ishida, Kana, Sakata, Rina C., Ichiraku, Minori, Takimoto, Ren, Yogo, Rina, Kijima, Yusuke, Mori, Hideto, Tanaka, Mamoru, King, Samuel, Tarumoto, Shoko, Tsujimura, Taro, Bashth, Omar, Masuyama, Nanami, Adel, Arman, Toyoshima, Hiromi, Seki, Motoaki, Oh, Ju Hee, Archambault, Anne-Sophie, Nishida, Keiji, Kondo, Akihiko, Kuhara, Satoru, Aburatani, Hiroyuki, Klein Geltink, Ramon I., Yamamoto, Takuya, Shakiba, Nika, Takashima, Yasuhiro, Yachie, Nozomu

Issue&Volume: 2025-05-21

Abstract: Cell-tagging strategies with DNA barcodes have enabled the analysis of clone size dynamics and clone-restricted transcriptomic landscapes in heterogeneous populations. However, isolating a target clone that displays a specific phenotype from a complex population remains challenging. Here we present a multi-kingdom genetic barcoding system, CloneSelect, which enables a target cell clone to be triggered to express a reporter gene for isolation through barcode-specific CRISPR base editing. In CloneSelect, cells are first stably tagged with DNA barcodes and propagated so that their subpopulation can be subjected to a given experiment. A clone that shows a phenotype or genotype of interest at a given time can then be isolated from the initial or subsequent cell pools stored during the experiment using CRISPR base editing. CloneSelect is scalable and compatible with single-cell RNA sequencing. We demonstrate the versatility of CloneSelect in human embryonic kidney 293T cells, mouse embryonic stem cells, human pluripotent stem cells, yeast cells and bacterial cells.

DOI: 10.1038/s41587-025-02649-1

Source: https://www.nature.com/articles/s41587-025-02649-1