Multiplex live single-cell transcriptional analysis demarcates cellular functional heterogeneity.
FRET
cell biology
cellular differentiation
cellular heterogeneity
developmental biology
human
lineage diversification
pluripotent stem cells
single-cell physiology
Journal
eLife
ISSN: 2050-084X
Titre abrégé: Elife
Pays: England
ID NLM: 101579614
Informations de publication
Date de publication:
08 10 2019
08 10 2019
Historique:
received:
23
06
2019
accepted:
07
10
2019
pubmed:
9
10
2019
medline:
9
4
2020
entrez:
9
10
2019
Statut:
epublish
Résumé
A fundamental goal in the biological sciences is to determine how individual cells with varied gene expression profiles and diverse functional characteristics contribute to development, physiology, and disease. Here, we report a novel strategy to assess gene expression and cell physiology in single living cells. Our approach utilizes fluorescently labeled mRNA-specific anti-sense RNA probes and dsRNA-binding protein to identify the expression of specific genes in real-time at single-cell resolution via FRET. We use this technology to identify distinct myocardial subpopulations expressing the structural proteins myosin heavy chain α and myosin light chain 2a in real-time during early differentiation of human pluripotent stem cells. We combine this live-cell gene expression analysis with detailed physiologic phenotyping to capture the functional evolution of these early myocardial subpopulations during lineage specification and diversification. This live-cell mRNA imaging approach will have wide ranging application wherever heterogeneity plays an important biological role.
Identifiants
pubmed: 31591966
doi: 10.7554/eLife.49599
pii: 49599
pmc: PMC6861004
doi:
pii:
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : NHLBI NIH HHS
ID : U01 HL100408
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL130391
Pays : United States
Organisme : American Heart Association
ID : Predoctoral Fellowship
Pays : International
Organisme : NHLBI NIH HHS
ID : Progenitor Cell Biology Consortium (PCBC) Jump Start Award
Pays : United States
Organisme : American Heart Association
ID : Predoctoral Fellowship: 15PRE22220009
Pays : International
Organisme : NHLBI NIH HHS
ID : Progenitor Cell Biology Consortium (PCBC) Jump Start Award: 5U01HL099997-07
Pays : United States
Organisme : NHLBI NIH HHS
ID : U01 HL099997
Pays : United States
Organisme : NHLBI NIH HHS
ID : U01HL100408-01
Pays : United States
Informations de copyright
© 2019, Atmanli et al.
Déclaration de conflit d'intérêts
AA, ID Inventor of a pending patent (PCT/US2016/029972), DH, FD, Av, FC, LB No competing interests declared
Références
J Cell Sci. 2010 May 15;123(Pt 10):1761-74
pubmed: 20427315
Biotechniques. 2004 Jan;36(1):114-22
pubmed: 14740493
Annu Rev Biomed Eng. 2009;11:25-47
pubmed: 19400712
Cardiovasc Res. 1997 Aug;35(2):315-23
pubmed: 9349394
Curr Opin Chem Biol. 2011 Dec;15(6):806-12
pubmed: 22055496
Nature. 2009 Jul 2;460(7251):113-7
pubmed: 19571884
Science. 1998 Apr 24;280(5363):585-90
pubmed: 9554849
Nat Biotechnol. 1996 Mar;14(3):303-8
pubmed: 9630890
Proc Natl Acad Sci U S A. 2015 Oct 13;112(41):12705-10
pubmed: 26417073
Development. 2015 Sep 15;142(18):3231-8
pubmed: 26209647
Circ J. 2013;77(5):1307-14
pubmed: 23400258
Stem Cells. 2008 Aug;26(8):1961-72
pubmed: 18483424
Nat Neurosci. 2013 Jul;16(7):780-9
pubmed: 23799470
Cell Stem Cell. 2018 Oct 4;23(4):586-598.e8
pubmed: 30290179
Nat Methods. 2007 May;4(5):413-9
pubmed: 17401370
Genom Data. 2015 Apr 01;4:69-72
pubmed: 26484180
Nat Protoc. 2013 Feb;8(2):265-81
pubmed: 23306460
Adv Healthc Mater. 2014 Nov;3(11):1759-64
pubmed: 24805162
Cell. 2016 Apr 7;165(2):488-96
pubmed: 26997482
RNA. 2008 Jun;14(6):1201-13
pubmed: 18426922
Biophys J. 2004 Dec;87(6):4153-62
pubmed: 15377515
Dev Cell. 2016 Nov 21;39(4):491-507
pubmed: 27840109
Cell Rep. 2017 Jan 17;18(3):777-790
pubmed: 28099854
Biophys J. 2010 Aug 9;99(4):1274-83
pubmed: 20713013
Cell. 2014 Oct 9;159(2):428-39
pubmed: 25303535
FEBS Lett. 2003 Jul 3;546(1):87-92
pubmed: 12829241
Dev Dyn. 2006 Jul;235(7):1994-2002
pubmed: 16649168
Dev Cell. 2016 Nov 21;39(4):480-490
pubmed: 27840107
J Clin Invest. 1996 Jul 1;98(1):216-24
pubmed: 8690796
Nat Biotechnol. 2017 Jan;35(1):56-68
pubmed: 27941801
Circ Res. 2014 Jan 31;114(3):511-23
pubmed: 24481842
Proc Natl Acad Sci U S A. 2012 Oct 23;109(43):17454-9
pubmed: 23064634
Proc Natl Acad Sci U S A. 1995 Oct 10;92(21):9445-9
pubmed: 7568151
Cell. 2006 Dec 15;127(6):1137-50
pubmed: 17123591
Cell. 2011 Nov 23;147(5):1054-65
pubmed: 22118462
Nat Methods. 2011 Oct 23;8(12):1037-40
pubmed: 22020065
Nat Methods. 2014 Aug;11(8):855-60
pubmed: 24930130
Cell Rep. 2015 Dec 29;13(12):2653-62
pubmed: 26711333
Stem Cell Res. 2013 Nov;11(3):1335-47
pubmed: 24095945
Nature. 2014 Dec 4;516(7529):56-61
pubmed: 25471879
Anat Rec. 1987 Sep;219(1):53-9
pubmed: 3688461
Nat Rev Drug Discov. 2012 Jan 20;11(2):125-40
pubmed: 22262036
Cell Stem Cell. 2015 Jun 4;16(6):712-24
pubmed: 26004780
J Cell Sci. 2009 Jun 15;122(Pt 12):2119-26
pubmed: 19470580
Biophys J. 2001 Oct;81(4):2395-402
pubmed: 11566809
Traffic. 2011 Aug;12(8):1000-11
pubmed: 21518164
Genes Cells. 2002 Sep;7(9):881-7
pubmed: 12296819
Nat Methods. 2011 Nov 06;8(12):1027-36
pubmed: 22056676
Mol Cell. 1998 Oct;2(4):437-45
pubmed: 9809065
RNA. 2003 Mar;9(3):364-74
pubmed: 12592010
Methods Cell Biol. 2013;114:593-609
pubmed: 23931524
Science. 2009 Oct 16;326(5951):426-9
pubmed: 19833966
Dev Cell. 2019 Feb 25;48(4):475-490.e7
pubmed: 30713072
J Cell Biol. 2015 May 25;209(4):609-19
pubmed: 26008747
Circulation. 2013 Oct 22;128(17):1897-909
pubmed: 23995537
Biomaterials. 2015 May;50:176-85
pubmed: 25736507
Biochemistry. 2012 Nov 20;51(46):9312-22
pubmed: 23140277
Cell Rep. 2019 Feb 12;26(7):1934-1950.e5
pubmed: 30759401
J Neurosci. 2016 Mar 30;36(13):3811-20
pubmed: 27030765
Nat Methods. 2009 May;6(5):347-9
pubmed: 19349979
Am J Physiol Heart Circ Physiol. 2004 Apr;286(4):H1471-9
pubmed: 14670817
Biochemistry. 1996 Aug 6;35(31):9983-94
pubmed: 8756460
Biochemistry. 2007 Jan 9;46(1):55-65
pubmed: 17198375
Science. 2014 Jan 24;343(6169):419-22
pubmed: 24458642
Cell Stem Cell. 2013 May 2;12(5):559-72
pubmed: 23642365
PLoS Biol. 2006 Oct;4(10):e309
pubmed: 17048983
Nat Methods. 2009 May;6(5):331-8
pubmed: 19404252
Cell Stem Cell. 2013 Jan 3;12(1):101-13
pubmed: 23290139
Genes Dev. 1992 Dec;6(12B):2478-90
pubmed: 1364113
Proc Natl Acad Sci U S A. 2012 Jul 3;109(27):E1848-57
pubmed: 22645348
Nature. 2013 Sep 19;501(7467):328-37
pubmed: 24048065
PLoS One. 2013 Apr 16;8(4):e61096
pubmed: 23613792