RNA imaging. Spatially resolved, highly multiplexed RNA profiling in single cells.
RNA imaging. Spatially resolved, highly multiplexed RNA profiling in single cells.
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DOI：
10.1126/science.aaa6090
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发表时间：
2015-04-24
期刊：
影响因子：
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通讯作者：
Zhuang X
中科院分区：
文献类型：
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作者：
Chen KH;Boettiger AN;Moffitt JR;Wang S;Zhuang X
了解单个细胞中转录组的表达谱和空间格局对于理解丰富的细胞行为库是必不可少的。在这里，我们报告多重错误鲁棒荧光原位杂交（MERFISH），一种单分子成像方法，允许在单细胞中确定数千种RNA的拷贝数和空间定位。使用错误稳健的编码方案来对抗单分子标记和检测错误，我们展示了数百个单个细胞中100 - 1000种独特RNA的成像。~104 - 106对基因的相关性分析使我们能够约束基因调控网络，预测许多未注释基因的新功能，并确定与编码蛋白质性质相关的RNA的不同空间分布模式。
Knowledge of the expression profile and spatial landscape of the transcriptome in individual cells is essential for understanding the rich repertoire of cellular behaviors. Here we report multiplexed error-robust fluorescence in situ hybridization (MERFISH), a single-molecule imaging approach that allows the copy numbers and spatial localizations of thousands of RNA species to be determined in single cells. Using error-robust encoding schemes to combat single-molecule labeling and detection errors, we demonstrated the imaging of 100 – 1000 unique RNA species in hundreds of individual cells. Correlation analysis of the ~104 – 106 pairs of genes allowed us to constrain gene regulatory networks, predict novel functions for many unannotated genes, and identify distinct spatial distribution patterns of RNAs that correlate with properties of the encoded proteins.