A multi-kingdom genetic barcoding system for precise target clone isolation
A multi-kingdom genetic barcoding system for precise target clone isolation
复制标题
DOI：
10.1101/2023.01.18.524633
复制
发表时间：
2023-01
期刊：
影响因子：
--
通讯作者：
Soh Ishiguro;Kana Ishida;Rina C. Sakata;Hideto Mori;Mamoru Takana;Samuel King;Omar S. Bashth;Minori Ichiraku;Nanami Masuyama;Ren Takimoto;Yusuke Kijima;Arman Adel;H. Toyoshima;Motoaki Seki;Ju Hee Oh;Anne-Sophie Archambault;K. Nishida;A. Kondo;S. Kuhara;H. Aburatani;R. K. Klein Geltink;Yasuhiro Takashima;Nika Shakiba;Nozomu Yachie
中科院分区：
文献类型：
--
作者：
Soh Ishiguro;Kana Ishida;Rina C. Sakata;Hideto Mori;Mamoru Takana;Samuel King;Omar S. Bashth;Minori Ichiraku;Nanami Masuyama;Ren Takimoto;Yusuke Kijima;Arman Adel;H. Toyoshima;Motoaki Seki;Ju Hee Oh;Anne-Sophie Archambault;K. Nishida;A. Kondo;S. Kuhara;H. Aburatani;R. K. Klein Geltink;Yasuhiro Takashima;Nika Shakiba;Nozomu Yachie
克隆异质性是多种生物学过程的基础，包括癌症进展、细胞分化和微生物进化。最近，使用DNA条形码的细胞标记策略使得能够分析异质群体的克隆大小动态和克隆限制性转录组学景观。然而，从复杂的群体中分离显示特定表型的靶克隆仍然具有挑战性。在这里，我们提出了一种新的多王国遗传条形码系统CloneSelect，其中可以通过条形码特异性CRISPR碱基编辑触发靶细胞克隆表达报告基因以进行分离。在CloneSelect中，细胞首先被条形码化并繁殖，因此它们的亚群可以进行给定的实验。然后可以从在整个实验时间过程中储存的初始或后续细胞库中分离在给定时间显示出感兴趣的表型或基因型的克隆。这种新型CRISPR条形码遗传学平台提供了许多分析和操纵哺乳动物、酵母和细菌系统的新方法。一种多王国CRISPR可激活的条形码系统能够从复杂的细胞群中精确分离目标条形码标记的克隆。
Clonal heterogeneity underlies diverse biological processes, including cancer progression, cell differentiation, and microbial evolution. Cell tagging strategies with DNA barcodes have recently enabled analysis of clone size dynamics and clone-restricted transcriptomic landscapes of heterogeneous populations. However, isolating a target clone that displays a specific phenotype from a complex population remains challenging. Here, we present a new multi-kingdom genetic barcoding system, CloneSelect, in which a target cell clone can be triggered to express a reporter gene for isolation through barcode-specific CRISPR base editing. In CloneSelect, cells are first barcoded and propagated so 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 throughout the experimental timecourse. This novel CRISPR-barcode genetics platform provides many new ways of analyzing and manipulating mammalian, yeast, and bacterial systems. Teaser A multi-kingdom CRISPR-activatable barcoding system enables the precise isolation of target barcode-labeled clones from a complex cell population.