SCITO-seq: single-cell combinatorial indexed cytometry sequencing.
SCITO-seq: single-cell combinatorial indexed cytometry sequencing.
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DOI：
10.1038/s41592-021-01222-3
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发表时间：
2021-08
期刊：
影响因子：
48
通讯作者：
Ye CJ
中科院分区：
文献类型：
--
作者：
Hwang B;Lee DS;Tamaki W;Sun Y;Ogorodnikov A;Hartoularos GC;Winters A;Yeung BZ;Nazor KL;Song YS;Chow ED;Spitzer MH;Ye CJ
开发DNA条形码抗体来标记细胞表面分子，使得能够使用基于液滴的单细胞测序（dsc-seq）来同时分析数千个细胞的蛋白质丰度。与流式细胞术和质谱仪相比，目前基于dsc-seq的工作流程的高单位细胞成本阻碍了其在临床应用和大规模合并筛选中的使用。在这里，我们介绍了SCITO-seq，这是一种使用夹板寡核苷酸（oligo）的工作流程，可以使用商业微流体技术对每个反应超过105个细胞的DNA条形码抗体进行组合索引dsc-seq。通过将样品条形码编码到夹板寡核苷酸中，我们证明了多重SCITO-seq产生与质谱细胞术相当的细胞组成和表面蛋白表达的可重复估计。我们进一步展示了两种修饰的夹板寡核苷酸设计，其扩展了SCITO-seq以实现与商业DNA条形码化抗体的相容性，并同时表达来自同一细胞的转录组和表面蛋白。这些结果表明，SCITO-seq是一种灵活的超高通量平台，可用于基于测序的单细胞蛋白质和多模式分析。
The development of DNA-barcoded antibodies to tag cell-surface molecules has enabled the use of droplet-based single cell sequencing (dsc-seq) to profile protein abundances from thousands of cells simultaneously. Compared to flow and mass cytometry, high per cell cost of current dsc-seq-based workflows precludes their use in clinical applications and large-scale pooled screens. Here, we introduce SCITO-seq, a workflow that uses splint oligonucleotides (oligos) to enable combinatorially indexed dsc-seq of DNA-barcoded antibodies from greater than 105 cells per reaction using commercial microfluidics. By encoding sample barcodes into splint oligos, we demonstrate that multiplexed SCITO-seq produces reproducible estimates of cellular composition and surface protein expression comparable to mass cytometry. We further demonstrate two modified splint oligo designs that extend SCITO-seq to achieve compatibility with commercial DNA-barcoded antibodies and simultaneous expression profiling of the transcriptome and surface proteins from the same cell. These results demonstrate SCITO-seq as a flexible and ultra high-throughput platform for sequencing-based single-cell protein and multimodal profiling.
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影响因子：
64.8
作者：
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通讯作者：
Shendure, Jay
影响因子：
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作者：
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作者：
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通讯作者：
Bielas JH