Multiplexed labeling of genomic loci with dCas9 and engineered sgRNAs using CRISPRainbow.
Multiplexed labeling of genomic loci with dCas9 and engineered sgRNAs using CRISPRainbow.
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DOI：
10.1038/nbt.3526
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发表时间：
2016-05
影响因子：
46.9
通讯作者：
Pederson T
中科院分区：
文献类型：
--
作者：
Ma H;Tu LC;Naseri A;Huisman M;Zhang S;Grunwald D;Pederson T
缺乏在活细胞中成像多个基因组位点的技术限制了我们研究染色体动力学的能力。在这里，我们描述了CRISPRainbow，一种基于核酸酶死亡（d）Cas9结合结合结合荧光蛋白组的工程化单向导RNA（sgRNA）支架的活细胞中标记DNA的系统。我们展示了在单个活细胞中多达六个染色体位点的同时成像，并记录了不同染色体位点的动态特性的巨大差异。
A lack of techniques to image multiple genomic loci in living cells has limited our ability to investigate chromosome dynamics. Here we describe CRISPRainbow, a system for labeling DNA in living cells based on nuclease-dead (d) Cas9 combined with engineered single guide RNA (sgRNA) scaffolds that bind sets of fluorescent proteins. We demonstrate simultaneous imaging of up to six chromosomal loci in individual live cells and document large differences in the dynamic properties of different chromosomal loci.