小柯机器人

美国加州大学旧金山分校Chun Jimmie Ye团队开发出单细胞组合索引细胞测序法。该研究于2021年8月5日发表于国际一流学术期刊《自然—方法学》。

研究人员报道了SCITO-seq，这是一个使用拼接寡核苷酸（oligos）的工作流程，可以使用商业微流控技术对每个反应中超过10⁵个细胞的DNA编码抗体进行组合索引的dsc-seq。通过将样品条形码编码到夹层寡头中，研究人员证明了多重SCITO-seq产生的细胞组成和表面蛋白表达的可重复性估计可与质谱法相媲美。研究人员进一步展示了两种改良的夹层寡头设计，扩展了SCITO-seq，从而实现与商业DNA编码抗体的兼容，并同时对同一细胞的转录组和表面蛋白进行表达分析。这些结果表明，SCITO-seq是一个灵活的、超高通量的平台，可用于基于测序的单细胞蛋白质和多模态分析。

据了解，标记细胞表面分子的DNA编码抗体的发展使得基于液滴的单细胞测序（dsc-seq）能够同时对成千上万的细胞进行蛋白质丰度分析。与流式细胞仪和质量细胞仪相比，目前基于dsc-seq的工作流程的每个细胞成本很高，这使得它们无法用于临床应用和大规模的集合筛选。

附：英文原文

Title: SCITO-seq: single-cell combinatorial indexed cytometry sequencing

Author: Hwang, Byungjin, Lee, David S., Tamaki, Whitney, Sun, Yang, Ogorodnikov, Anton, Hartoularos, George C., Winters, Aidan, Yeung, Bertrand Z., Nazor, Kristopher L., Song, Yun S., Chow, Eric D., Spitzer, Matthew H., Ye, Chun Jimmie

Issue&Volume: 2021-08-05

Abstract: 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. As compared to flow and mass cytometry, the 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 over 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 those from 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. 

DOI: 10.1038/s41592-021-01222-3

Source: https://www.nature.com/articles/s41592-021-01222-3

Nature Methods：《自然—方法学》，创刊于2004年。隶属于施普林格·自然出版集团，最新IF：47.99
官方网址：https://www.nature.com/nmeth/
投稿链接：https://mts-nmeth.nature.com/cgi-bin/main.plex