Multibow: digital spectral barcodes for cell tracing.

Multibow: digital spectral barcodes for cell tracing.
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DOI:
10.1371/journal.pone.0127822
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发表时间:
2015
期刊:
影响因子:
3.7
通讯作者:
Megason SG
Megason SG
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Xiong F;Obholzer ND;Noche RR;Megason SG

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我们介绍了一种用于发育和再生过程中细胞示踪实验的标记策略(Multibow)。基于Brainbow的方法,通过不同荧光蛋白的差异表达水平产生颜色,Multibow通过引入二进制代码增加了一层标签多样性,其中报告基因最初为OFF,然后在Cre重组后概率性地ON或OFF。我们已经开发了一个包含七种不同颜色和三种不同亚细胞定位的结构库。在Cre的存在下组合来自该文库的构建体产生用多种独立表达的颜色标记的细胞,所述颜色基于每个构建体在重组后是ON还是OFF。这些标记形成独特的“条形码”,除了每种颜色的表达水平差异之外,还允许跟踪细胞及其克隆后代。我们在斑马鱼中测试了Multibow,验证了其设计理念,并建议其在发育和再生中的细胞追踪应用程序的实用性。
We introduce a multicolor labeling strategy (Multibow) for cell tracing experiments in developmental and regenerative processes. Building on Brainbow-based approaches that produce colors by differential expression levels of different fluorescent proteins, Multibow adds a layer of label diversity by introducing a binary code in which reporters are initially OFF and then probabilistically ON or OFF following Cre recombination. We have developed a library of constructs that contains seven different colors and three different subcellular localizations. Combining constructs from this library in the presence of Cre generates cells labeled with multiple independently expressed colors based on if each construct is ON or OFF following recombination. These labels form a unique "barcode" that allows the tracking of the cell and its clonal progenies in addition to expression level differences of each color. We tested Multibow in zebrafish which validates its design concept and suggests its utility for cell tracing applications in development and regeneration.
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