PT  - JOURNAL ARTICLE
AU  - Luke Vistain
AU  - Hoang Van Phan
AU  - Christian Jordi
AU  - Mengjie Chen
AU  - Sai T. Reddy
AU  - Savaş Tay
TI  - Quantification of proteins, protein complexes and mRNA in single cells by proximity-sequencing
AID  - 10.1101/2020.05.15.098780
DP  - 2020 Jan 01
TA  - bioRxiv
PG  - 2020.05.15.098780
4099  - http://biorxiv.org/content/early/2020/05/16/2020.05.15.098780.short
4100  - http://biorxiv.org/content/early/2020/05/16/2020.05.15.098780.full
AB  - 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 cell’s 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.Competing Interest StatementThe authors have declared no competing interest.