A programmable sequence of reporters for lineage analysis
A programmable sequence of reporters for lineage analysis
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DOI：
10.1038/s41593-020-0676-9
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发表时间：
2020-07-27
影响因子：
25
通讯作者：
Lee, Tzumin
中科院分区：
文献类型：
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作者：
Garcia-Marques, Jorge;Espinosa-Medina, Isabel;Lee, Tzumin
Garcia-Marques等人提出了CLADES，这是一种基于CRISPR研究神经元谱系的创新方法。受合成生物学的启发，CLADES依赖于一个基因开关系统，以预定的顺序激活和激活报告基因。我们介绍了CLADES（由编辑序列驱动的细胞谱系访问），一种基于CRISPR-Cas9技术的细胞谱系研究技术。CLADES依赖于一个基因开关系统，以预定的顺序激活和激活报告基因。将CLADES靶向祖细胞允许后代继承报告基因的连续级联，从而将出生顺序与报告基因表达偶联。该系统也可以通过热休克暂时诱导，使得谱系发育的时间分辨率成为可能，因此可以通过追踪子代中表达的报告基因来解构扩展的细胞谱系。当靶向生殖系时，相同的级联在动物代间进行，主要用相应的报告基因组合标记每一代。因此，CLADES提供了一种创新的策略，使可编程的基因级联，可用于遗传操作或记录一系列生物事件。
Garcia-Marques et al. present CLADES, an innovative approach to study neuronal lineages based on CRISPR. Inspired by synthetic biology, CLADES relies on a system of genetic switches to activate and inactivate reporter genes in a predetermined order.We present CLADES (cell lineage access driven by an edition sequence), a technology for cell lineage studies based on CRISPR-Cas9 techniques. CLADES relies on a system of genetic switches to activate and inactivate reporter genes in a predetermined order. Targeting CLADES to progenitor cells allows the progeny to inherit a sequential cascade of reporters, thereby coupling birth order to reporter expression. This system, which can also be temporally induced by heat shock, enables the temporal resolution of lineage development and can therefore be used to deconstruct an extended cell lineage by tracking the reporters expressed in the progeny. When targeted to the germ line, the same cascade progresses across animal generations, predominantly marking each generation with the corresponding combination of reporters. CLADES therefore offers an innovative strategy for making programmable cascades of genes that can be used for genetic manipulation or to record serial biological events.