Highly multiplexed simultaneous detection of RNAs and proteins in single cells.

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.

journal_name

Nat Methods

journal_title

Nature methods

authors

Frei AP,Bava FA,Zunder ER,Hsieh EW,Chen SY,Nolan GP,Gherardini PF

doi

10.1038/nmeth.3742

subject

Has Abstract

pub_date

2016-03-01 00:00:00

pages

269-75

issue

3

eissn

1548-7091

issn

1548-7105

pii

nmeth.3742

journal_volume

13

pub_type

杂志文章
  • Atomic-accuracy models from 4.5-Å cryo-electron microscopy data with density-guided iterative local refinement.

    abstract::We describe a general approach for refining protein structure models on the basis of cryo-electron microscopy maps with near-atomic resolution. The method integrates Monte Carlo sampling with local density-guided optimization, Rosetta all-atom refinement and real-space B-factor fitting. In tests on experimental maps o...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth.3286

    authors: DiMaio F,Song Y,Li X,Brunner MJ,Xu C,Conticello V,Egelman E,Marlovits T,Cheng Y,Baker D

    更新日期:2015-04-01 00:00:00

  • Whole-genome genotyping with the single-base extension assay.

    abstract::We describe an efficient, accurate and robust whole-genome genotyping (WGG) assay based on a two-color, single-base extension (SBE), single-nucleotide polymorphism (SNP)-scoring step. We report genotyping results for biallelic International HapMap quality control (QC) SNPs using a single probe per locus. We show scala...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth842

    authors: Steemers FJ,Chang W,Lee G,Barker DL,Shen R,Gunderson KL

    更新日期:2006-01-01 00:00:00

  • Single-cell genomics.

    abstract::Methods for genomic analysis at single-cell resolution enable new understanding of complex biological phenomena. Single-cell techniques, ranging from flow cytometry and microfluidics to PCR and sequencing, are used to understand the cellular composition of complex tissues, find new microbial species and perform genome...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth0411-311

    authors: Kalisky T,Quake SR

    更新日期:2011-04-01 00:00:00

  • Measurement of mechanical tractions exerted by cells in three-dimensional matrices.

    abstract::Quantitative measurements of cell-generated forces have heretofore required that cells be cultured on two-dimensional substrates. We describe a technique to quantitatively measure three-dimensional traction forces exerted by cells fully encapsulated in well-defined elastic hydrogel matrices. Using this approach we mea...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth.1531

    authors: Legant WR,Miller JS,Blakely BL,Cohen DM,Genin GM,Chen CS

    更新日期:2010-12-01 00:00:00

  • A streamlined platform for high-content functional proteomics of primary human specimens.

    abstract::Achieving information content of satisfactory breadth and depth remains a formidable challenge for proteomics. This problem is particularly relevant to the study of primary human specimens, such as tumor biopsies, which are heterogeneous and of finite quantity. Here we present a functional proteomics strategy that uni...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth778

    authors: Jessani N,Niessen S,Wei BQ,Nicolau M,Humphrey M,Ji Y,Han W,Noh DY,Yates JR 3rd,Jeffrey SS,Cravatt BF

    更新日期:2005-09-01 00:00:00

  • High-frequency genome editing using ssDNA oligonucleotides with zinc-finger nucleases.

    abstract::Zinc-finger nucleases (ZFNs) have enabled highly efficient gene targeting in multiple cell types and organisms. Here we describe methods for using simple ssDNA oligonucleotides in tandem with ZFNs to efficiently produce human cell lines with three distinct genetic outcomes: (i) targeted point mutation, (ii) targeted g...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth.1653

    authors: Chen F,Pruett-Miller SM,Huang Y,Gjoka M,Duda K,Taunton J,Collingwood TN,Frodin M,Davis GD

    更新日期:2011-07-17 00:00:00

  • Designer receptors for every body.

    abstract::Using a directed evolution approach, researchers demonstrate a way of creating 'designer' receptors that are specifically activated by a ligand with no other biological activity in the cell. ...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth0507-382b

    authors: Kaganman I

    更新日期:2007-05-01 00:00:00

  • Optimized ratiometric calcium sensors for functional in vivo imaging of neurons and T lymphocytes.

    abstract::The quality of genetically encoded calcium indicators (GECIs) has improved dramatically in recent years, but high-performing ratiometric indicators are still rare. Here we describe a series of fluorescence resonance energy transfer (FRET)-based calcium biosensors with a reduced number of calcium binding sites per sens...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth.2773

    authors: Thestrup T,Litzlbauer J,Bartholomäus I,Mues M,Russo L,Dana H,Kovalchuk Y,Liang Y,Kalamakis G,Laukat Y,Becker S,Witte G,Geiger A,Allen T,Rome LC,Chen TW,Kim DS,Garaschuk O,Griesinger C,Griesbeck O

    更新日期:2014-02-01 00:00:00

  • Strategy for the fine characterization of glycosyltransferase specificity using isotopomer assembly.

    abstract::Glycosylation, which represents the most complex posttranslational modification (PTM) event during protein maturation, has a vital role in biological processes. Glycan biosynthesis is orchestrated by numerous glycosyltransferases, each displaying different selectivities for multiple reaction sites. The precise specifi...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth1050

    authors: Ito H,Kameyama A,Sato T,Sukegawa M,Ishida HK,Narimatsu H

    更新日期:2007-07-01 00:00:00

  • Conformal nanopatterning of extracellular matrix proteins onto topographically complex surfaces.

    abstract::Our Patterning on Topography (PoT) printing technique enables fibronectin, laminin and other proteins to be applied to biomaterial surfaces in complex geometries that are inaccessible using traditional soft lithography techniques. Engineering combinatorial surfaces that integrate topographical and biochemical micropat...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth.3210

    authors: Sun Y,Jallerat Q,Szymanski JM,Feinberg AW

    更新日期:2015-02-01 00:00:00

  • fMRIPrep: a robust preprocessing pipeline for functional MRI.

    abstract::Preprocessing of functional magnetic resonance imaging (fMRI) involves numerous steps to clean and standardize the data before statistical analysis. Generally, researchers create ad hoc preprocessing workflows for each dataset, building upon a large inventory of available tools. The complexity of these workflows has s...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/s41592-018-0235-4

    authors: Esteban O,Markiewicz CJ,Blair RW,Moodie CA,Isik AI,Erramuzpe A,Kent JD,Goncalves M,DuPre E,Snyder M,Oya H,Ghosh SS,Wright J,Durnez J,Poldrack RA,Gorgolewski KJ

    更新日期:2019-01-01 00:00:00

  • Nanoscale resolution in GFP-based microscopy.

    abstract::We report attainment of subdiffraction resolution using stimulated emission depletion (STED) microscopy with GFP-labeled samples. The approximately 70 nm lateral resolution attained in this study is demonstrated by imaging GFP-labeled viruses and the endoplasmic reticulum (ER) of a mammalian cell. Our results mark the...

    journal_title:Nature methods

    pub_type: 杂志文章,评审

    doi:10.1038/nmeth922

    authors: Willig KI,Kellner RR,Medda R,Hein B,Jakobs S,Hell SW

    更新日期:2006-09-01 00:00:00

  • Prosit: proteome-wide prediction of peptide tandem mass spectra by deep learning.

    abstract::In mass-spectrometry-based proteomics, the identification and quantification of peptides and proteins heavily rely on sequence database searching or spectral library matching. The lack of accurate predictive models for fragment ion intensities impairs the realization of the full potential of these approaches. Here, we...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/s41592-019-0426-7

    authors: Gessulat S,Schmidt T,Zolg DP,Samaras P,Schnatbaum K,Zerweck J,Knaute T,Rechenberger J,Delanghe B,Huhmer A,Reimer U,Ehrlich HC,Aiche S,Kuster B,Wilhelm M

    更新日期:2019-06-01 00:00:00

  • Functional ultrasound imaging of the brain.

    abstract::We present functional ultrasound (fUS), a method for imaging transient changes in blood volume in the whole brain at better spatiotemporal resolution than with other functional brain imaging modalities. fUS uses plane-wave illumination at high frame rate and can measure blood volumes in smaller vessels than previous u...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth.1641

    authors: Macé E,Montaldo G,Cohen I,Baulac M,Fink M,Tanter M

    更新日期:2011-07-03 00:00:00

  • Cas9 as a versatile tool for engineering biology.

    abstract::RNA-guided Cas9 nucleases derived from clustered regularly interspaced short palindromic repeats (CRISPR)-Cas systems have dramatically transformed our ability to edit the genomes of diverse organisms. We believe tools and techniques based on Cas9, a single unifying factor capable of colocalizing RNA, DNA and protein,...

    journal_title:Nature methods

    pub_type: 杂志文章,评审

    doi:10.1038/nmeth.2649

    authors: Mali P,Esvelt KM,Church GM

    更新日期:2013-10-01 00:00:00

  • Live-cell super-resolved PAINT imaging of piconewton cellular traction forces.

    abstract::Despite the vital role of mechanical forces in biology, it still remains a challenge to image cellular force with sub-100-nm resolution. Here, we present tension points accumulation for imaging in nanoscale topography (tPAINT), integrating molecular tension probes with the DNA points accumulation for imaging in nanosc...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/s41592-020-0929-2

    authors: Brockman JM,Su H,Blanchard AT,Duan Y,Meyer T,Quach ME,Glazier R,Bazrafshan A,Bender RL,Kellner AV,Ogasawara H,Ma R,Schueder F,Petrich BG,Jungmann R,Li R,Mattheyses AL,Ke Y,Salaita K

    更新日期:2020-10-01 00:00:00

  • Targeted chromosome elimination from ES-somatic hybrid cells.

    abstract::To engineer a stem cell genome, we developed a technology for targeted elimination of chromosomes from mouse embryonic stem (ES)-somatic hybrid cells. Here we demonstrate the use of a universal chromosome elimination cassette (CEC) for elimination of a single embryonic stem cell (ESC)-derived chromosome 11 or 12, and ...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth973

    authors: Matsumura H,Tada M,Otsuji T,Yasuchika K,Nakatsuji N,Surani A,Tada T

    更新日期:2007-01-01 00:00:00

  • Dynamic characterization of growth and gene expression using high-throughput automated flow cytometry.

    abstract::Cells adjust to changes in environmental conditions using complex regulatory programs. These cellular programs are the result of an intricate interplay between gene expression, cellular growth and protein degradation. Technologies that enable simultaneous and time-resolved measurements of these variables are necessary...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth.2879

    authors: Zuleta IA,Aranda-Díaz A,Li H,El-Samad H

    更新日期:2014-04-01 00:00:00

  • Measurement of atom resolvability in cryo-EM maps with Q-scores.

    abstract::Cryogenic electron microscopy (cryo-EM) maps are now at the point where resolvability of individual atoms can be achieved. However, resolvability is not necessarily uniform throughout the map. We introduce a quantitative parameter to characterize the resolvability of individual atoms in cryo-EM maps, the map Q-score. ...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/s41592-020-0731-1

    authors: Pintilie G,Zhang K,Su Z,Li S,Schmid MF,Chiu W

    更新日期:2020-03-01 00:00:00

  • Large-scale electrophysiology with polymer-based electrodes.

    abstract:: ...

    journal_title:Nature methods

    pub_type: 评论,杂志文章

    doi:10.1038/s41592-019-0315-0

    authors: Vogt N

    更新日期:2019-02-01 00:00:00

  • A photoprotection strategy for microsecond-resolution single-molecule fluorescence spectroscopy.

    abstract::Time resolution of current single-molecule fluorescence techniques is limited to milliseconds because of dye blinking and bleaching. Here we introduce a photoprotection strategy that affords microsecond resolution by combining efficient triplet quenching by oxygen and Trolox with minimized bleaching via the oxygen rad...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth.1553

    authors: Campos LA,Liu J,Wang X,Ramanathan R,English DS,Muñoz V

    更新日期:2011-02-01 00:00:00

  • Post-translational selective intracellular silencing of acetylated proteins with de novo selected intrabodies.

    abstract::The ability to selectively interfere with post-translationally modified proteins would have many biological and therapeutic applications. However, post-translational modifications cannot be selectively targeted by nucleic-acid-based interference approaches. Here we describe post-translational intracellular silencing a...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth.4144

    authors: Chirichella M,Lisi S,Fantini M,Goracci M,Calvello M,Brandi R,Arisi I,D'Onofrio M,Di Primio C,Cattaneo A

    更新日期:2017-03-01 00:00:00

  • Onco-proteogenomics: cancer proteomics joins forces with genomics.

    abstract::The complexities of tumor genomes are rapidly being uncovered, but how they are regulated into functional proteomes remains poorly understood. Standard proteomics workflows use databases of known proteins, but these databases do not capture the uniqueness of the cancer transcriptome, with its point mutations, unusual ...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth.3138

    authors: Alfaro JA,Sinha A,Kislinger T,Boutros PC

    更新日期:2014-11-01 00:00:00

  • Bright monomeric red fluorescent protein with an extended fluorescence lifetime.

    abstract::Fluorescent proteins have become extremely popular tools for in vivo imaging and especially for the study of localization, motility and interaction of proteins in living cells. Here we report TagRFP, a monomeric red fluorescent protein, which is characterized by high brightness, complete chromophore maturation, prolon...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth1062

    authors: Merzlyak EM,Goedhart J,Shcherbo D,Bulina ME,Shcheglov AS,Fradkov AF,Gaintzeva A,Lukyanov KA,Lukyanov S,Gadella TW,Chudakov DM

    更新日期:2007-07-01 00:00:00

  • Imaging cellular ultrastructures using expansion microscopy (U-ExM).

    abstract::Determining the structure and composition of macromolecular assemblies is a major challenge in biology. Here we describe ultrastructure expansion microscopy (U-ExM), an extension of expansion microscopy that allows the visualization of preserved ultrastructures by optical microscopy. This method allows for near-native...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/s41592-018-0238-1

    authors: Gambarotto D,Zwettler FU,Le Guennec M,Schmidt-Cernohorska M,Fortun D,Borgers S,Heine J,Schloetel JG,Reuss M,Unser M,Boyden ES,Sauer M,Hamel V,Guichard P

    更新日期:2019-01-01 00:00:00

  • Segregation of molecules at cell division reveals native protein localization.

    abstract::We introduce a nonintrusive method exploiting single-cell variability after cell division to validate protein localization. We found that Clp proteases, widely reported to form biologically relevant foci, were uniformly distributed in Escherichia coli cells, and that many commonly used fluorescent proteins caused seve...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth.1955

    authors: Landgraf D,Okumus B,Chien P,Baker TA,Paulsson J

    更新日期:2012-04-08 00:00:00

  • Quantifying small numbers of antibodies with a 'near-universal' protein-DNA chimera.

    abstract::We present general means to greatly increase the sensitivity of antibody-based assays. Augmentation relies on a 'tadpole' protein-DNA chimera whose protein moiety binds most classes of mammalian antibodies but not avian immunoglobulin Y (IgY). We used this tadpole in affinity capture assays followed by real-time PCR t...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth1127

    authors: Burbulis I,Yamaguchi K,Yu R,Resnekov O,Brent R

    更新日期:2007-12-01 00:00:00

  • Localization-based super-resolution imaging meets high-content screening.

    abstract::Single-molecule localization microscopy techniques have proven to be essential tools for quantitatively monitoring biological processes at unprecedented spatial resolution. However, these techniques are very low throughput and are not yet compatible with fully automated, multiparametric cellular assays. This shortcomi...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth.4486

    authors: Beghin A,Kechkar A,Butler C,Levet F,Cabillic M,Rossier O,Giannone G,Galland R,Choquet D,Sibarita JB

    更新日期:2017-12-01 00:00:00

  • The GEM mapper: fast, accurate and versatile alignment by filtration.

    abstract::Because of ever-increasing throughput requirements of sequencing data, most existing short-read aligners have been designed to focus on speed at the expense of accuracy. The Genome Multitool (GEM) mapper can leverage string matching by filtration to search the alignment space more efficiently, simultaneously deliverin...

    journal_title:Nature methods

    pub_type: 杂志文章

    doi:10.1038/nmeth.2221

    authors: Marco-Sola S,Sammeth M,Guigó R,Ribeca P

    更新日期:2012-12-01 00:00:00

  • An FDR metric for top-down proteomics.

    abstract:: ...

    journal_title:Nature methods

    pub_type: 评论,杂志文章

    doi:10.1038/s41592-019-0343-9

    authors: Doerr A

    更新日期:2019-03-01 00:00:00