Multiplexed, targeted profiling of single-cell proteomes and transcriptomes in a single reaction
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13059_2016_Article_1045.pdf
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1.59 MB
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Checksum (MD5)
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Author(s)
Li, Shuqiang
Darmanis, Spyros
Lundberg, Martin
Fredriksson, Simon
Landegren, Ulf
Gallant, Caroline J.
Livak, Kenneth J.
Genshaft, Alex S.
Prakadan, Sanjay
Ziegler, Carly
Date Issued
September 2016
Journal
Genome Biology
Publisher
BioMed Central
Citation
Genshaft, Alex S et al. “Multiplexed, Targeted Profiling of Single-Cell Proteomes and Transcriptomes in a Single Reaction.” Genome Biology 17.1 (2016): n. pag.
Version
Final published version
Abstract
We present a scalable, integrated strategy for coupled protein and RNA detection from single cells. Our approach leverages the DNA polymerase activity of reverse transcriptase to simultaneously perform proximity extension assays and complementary DNA synthesis in the same reaction. Using the Fluidigm C1™ system, we profile the transcriptomic and proteomic response of a human breast adenocarcinoma cell line to a chemical perturbation, benchmarking against in situ hybridizations and immunofluorescence staining, as well as recombinant proteins, ERCC Spike-Ins, and population lysate dilutions. Through supervised and unsupervised analyses, we demonstrate synergies enabled by simultaneous measurement of single-cell protein and RNA abundances. Collectively, our generalizable approach highlights the potential for molecular metadata to inform highly-multiplexed single-cell analyses.
MIT Department
Massachusetts Institute of Technology. Institute for Medical Engineering & Science
Harvard University--MIT Division of Health Sciences and Technology
Massachusetts Institute of Technology. Department of Biology
Massachusetts Institute of Technology. Department of Chemistry
Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science
Ragon Institute of MGH, MIT and Harvard
Koch Institute for Integrative Cancer Research at MIT
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Creative Commons Attribution
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DOI of Published Version
https://doi.org/10.1186/s13059-016-1045-6
