Publication:
Digital Quantification of Proteins and mRNA in Single Mammalian Cells

dc.contributor.authorAlbayrak, CEM
dc.contributor.authorJordi, Christian A.
dc.contributor.authorZechner, Christoph
dc.contributor.authorLin, Jing
dc.contributor.authorBichsel, Colette A.
dc.contributor.authorKhammash, Mustafa
dc.contributor.authorTay, Savas
dc.contributor.institutionauthorALBAYRAK, CEM
dc.date.accessioned2021-05-17T20:59:08Z
dc.date.available2021-05-17T20:59:08Z
dc.date.issued2016-03-01T00:00:00Z
dc.description.abstractAbsolute quantification of macromolecules in single cells is critical for understanding and modeling biological systems that feature cellular heterogeneity. Here we show extremely sensitive and absolute quantification of both proteins and mRNA in single mammalian cells by a very practical workflow that combines proximity ligation assay (PLA) and digital PCR. This digital PLA method has femtomolar sensitivity, which enables the quantification of very small protein concentration changes over its entire 3-log dynamic range, a quality necessary for accounting for single-cell heterogeneity. We counted both endogenous (CD147) and exogenously expressed (GFP-p65) proteins from hundreds of single cells and determined the correlation between CD147 mRNA and the protein it encodes. Using our data, a stochastic two-state model of the central dogma was constructed and verified using joint mRNA/protein distributions, allowing us to estimate transcription burst sizes and extrinsic noise strength and calculate the transcription and translation rate constants in single mammalian cells.
dc.identifier.citationAlbayrak C., Jordi C. A. , Zechner C., Lin J., Bichsel C. A. , Khammash M., Tay S., -Digital Quantification of Proteins and mRNA in Single Mammalian Cells-, MOLECULAR CELL, cilt.61, sa.6, ss.914-924, 2016
dc.identifier.doi10.1016/j.molcel.2016.02.030
dc.identifier.scopus84962564367
dc.identifier.urihttp://hdl.handle.net/20.500.12645/28881
dc.identifier.wosWOS:000372327100014
dc.rightsinfo:eu-repo/semantics/openAccess
dc.titleDigital Quantification of Proteins and mRNA in Single Mammalian Cells
dc.typeArticle
dspace.entity.typePublication
local.avesis.id296c683b-bbd4-4ea3-a5dc-dc7533d980da
local.publication.isinternational1
relation.isAuthorOfPublicationced2804c-b9e5-4f25-9926-74404ade4f49
relation.isAuthorOfPublication.latestForDiscoveryced2804c-b9e5-4f25-9926-74404ade4f49
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