Simultaneous Multiplexed Measurement of RNA and Proteins in Single Cells

Significant advances have been made in methods to analyze genomes and transcriptomes of single cells, but to fully define cell states, proteins must also be accessed as central actors defining a cell’s phenotype. Methods currently used to analyze endogenous protein expression in single cells are lim...

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Bibliographic Details
Published in:Cell reports (Cambridge) 2016-01, Vol.14 (2), p.380-389
Main Authors: Darmanis, Spyros, Gallant, Caroline Julie, Marinescu, Voichita Dana, Niklasson, Mia, Segerman, Anna, Flamourakis, Georgios, Fredriksson, Simon, Assarsson, Erika, Lundberg, Martin, Nelander, Sven, Westermark, Bengt, Landegren, Ulf
Format: Article
Language:English
Subjects:
Cell Line, Tumor
Humans
Proteins - genetics
Proteomics - methods
Resource
RNA - genetics
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Summary:Significant advances have been made in methods to analyze genomes and transcriptomes of single cells, but to fully define cell states, proteins must also be accessed as central actors defining a cell’s phenotype. Methods currently used to analyze endogenous protein expression in single cells are limited in specificity, throughput, or multiplex capability. Here, we present an approach to simultaneously and specifically interrogate large sets of protein and RNA targets in lysates from individual cells, enabling investigations of cell functions and responses. We applied our method to investigate the effects of BMP4, an experimental therapeutic agent, on early-passage glioblastoma cell cultures. We uncovered significant heterogeneity in responses to treatment at levels of RNA and protein, with a subset of cells reacting in a distinct manner to BMP4. Moreover, we found overall poor correlation between protein and RNA at the level of single cells, with proteins more accurately defining responses to treatment. [Display omitted] •Multiplexed protein and RNA measurement in single cells•Distinct information for defining cell states from RNA and protein data•Heterogeneity of responses to the putative therapeutic BMP4 in glioblastoma cells Darmanis et al. present an approach to simultaneously measure levels of up to 96 transcripts and proteins in single cells. They apply this technique to study responses of glioblastoma cells to BMP4, a proposed therapeutic agent, and reveal heterogeneous responses and cell states.
ISSN:2211-1247
2211-1247
DOI:10.1016/j.celrep.2015.12.021