Cited by 343Single-cell and spatial transcriptomicsAdvanced Proteomics Techniques and ApplicationsBiotin and Related Studies

Highly multiplexed simultaneous detection of RNAs and proteins in single cells

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Frei, Andreas P, Bava, Felice-Alessio, Zunder, Eli R, Hsieh, Elena W Y, Chen, Shih-Yu, Nolan, Garry P, Gherardini, Pier Federico·2016·Nature Methods

Abstract

To enable the detection of expression signatures specific to individual cells, we developed PLAYR (proximity ligation assay for RNA), a method for highly multiplexed transcript quantification by flow and mass cytometry that is compatible with standard antibody staining. When used with mass cytometry, PLAYR allowed for the simultaneous quantification of more than 40 different mRNAs and proteins. In primary cells, we quantified multiple transcripts, with the identity and functional state of each analyzed cell defined on the basis of the expression of a separate set of transcripts or proteins. By expanding high-throughput deep phenotyping of cells beyond protein epitopes to include RNA expression, PLAYR opens a new avenue for the characterization of cellular metabolism.

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  • PLAYR simultaneously quantifies more than 40 targeted mRNAs and proteins from 10,000 single cells in several cell types.

    “For example, in 2016, the proximity ligation assay for RNA (PLAYR) was developed to simultaneously quantify more than 40 targeted mRNAs and proteins from 10,000 single cells in several cell types67.”

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Authors
Frei, A. P., Bava, F.-A., Zunder, E. R., Hsieh, E. W. Y., Chen, S.-Y., Nolan, G. P., & Gherardini, P. F.
Affiliations
  • Stanford University (US)
  • University of Virginia (US)
  • University of Colorado Denver (US)
Corresponding author
  • Nolan, Garry P
  • Gherardini, Pier Federico
Venue
Nature Methods
Published
2016-01-25
Volume
13
Issue
3
Pages
269-275
ISSN
  • 1548-7091 (ISSN-L)
  • 1548-7105
OpenAlex
W2253972942
Cited by
330
References
51
Citations by year
2026: 8 · 2025: 20 · 2024: 24 · 2023: 32 · 2022: 29 · 2021: 31 · 2020: 35 · 2019: 45 · 2018: 43 · 2017: 42 · 2016: 20
FWCI
25.98
Type
journal article
Language
en
Open access
Locations
  • Nature Methods (publishedVersion)
  • PubMed (publishedVersion)
  • Europe PMC (PubMed Central) (acceptedVersion)
  • PubMed Central (acceptedVersion)
Funders
  • National Institutes of Health (5r01ca196657-02, n01hv28183-13-0-0, 5r01hl120724-05)
  • National Heart, Lung, and Blood Institute (N01 HV028183, HHSN268201000034C, R01 - HL120724)
  • National Cancer Institute (R33CA183654, R21CA183660, R33CA183692, R01CA184968, 1R01CA19665701)
  • National Institute of General Medical Sciences (R01GM109836, 1R01GM10983601)
  • National Institute of Allergy and Infectious Diseases (U19AI100627, P01AI036535, U19AI057229, T32AI007328)
  • National Institute of Neurological Disorders and Stroke (1R01NS08953301, R01NS089533)
  • National Institute of Arthritis and Musculoskeletal and Skin Diseases (UH2AR067676)
  • Eunice Kennedy Shriver National Institute of Child Health and Human Development (K12HD000850)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Single-cell and spatial transcriptomics
  • Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Biotin and Related Studies
Keywords
single-cell transcriptomics, single-cell proteomics, mass cytometry, flow cytometry, cellular metabolism, deep phenotyping, primary cells
MeSH terms
Flow Cytometry, Humans, Proteins, RNA, Jurkat Cells, Protein Array Analysis, High-Throughput Screening Assays, High-Throughput Nucleotide Sequencing

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