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bioRxiv · 10.1101/2020.05.15.098780

Quantification of proteins, protein complexes and mRNA in single cells by proximity-sequencing

Vistain, L.·Phan, H. V.·Jordi, C.·Chen, M.·Reddy, S. T.·Tay, S.
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Abstract

Multiplexed analysis of single-cells enables accurate modeling of cellular behaviors, classification of new cell types, and characterization of their functional states. Here we present proximity-sequencing (Prox-seq), a method for simultaneous measurement of an individual cells proteins, protein complexes and mRNA. Prox-seq utilizes deep sequencing and barcoded proximity assays to measure proteins and their complexes from all pairwise combinations of targeted proteins, in thousands of single-cells. The number of measured protein complexes scales quadratically with the number of targeted proteins, providing unparalleled multiplexing capacity. We developed a high-throughput experimental and computational pipeline and demonstrated the potential of Prox-Seq for multi-omic analysis with a panel of 13 barcoded proximity probes, enabling the measurement of 91 protein complexes, along with thousands of mRNA molecules in single T-cells and B-cells. Prox-seq provides access to an untapped yet powerful measurement modality for single-cell phenotyping and can discover new protein interactions in signaling and drug studies.

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Vistain, L., Phan, H. V., Jordi, C., Chen, M., Reddy, S. T., Tay, S.. 2020-05-16. Quantification of proteins, protein complexes and mRNA in single cells by proximity-sequencing. https://doi.org/10.1101/2020.05.15.098780

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