Imaging individual mRNA molecules using multiple singly labeled probes

被引：1351

作者：

Raj, Arjun ^{[1
]}

van den Bogaard, Patrick ^{[2
]}

Rifkin, Scott A. ^{[1
]}

van Oudenaarden, Alexander ^{[1
]}

Tyagi, Sanjay ^{[2
]}

机构：

[1] MIT, Dept Phys, Cambridge, MA 02139 USA

[2] Univ Med & Dent New Jersey, New Jersey Med Sch, Publ Hlth Res Inst Ctr, Newark, NJ 07103 USA

来源：

NATURE METHODS | 2008年 / 5卷 / 10期

基金：

美国国家科学基金会;

关键词：

D O I：

10.1038/NMETH.1253

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We describe a method for imaging individual mRNA molecules in fixed cells by probing each mRNA species with 48 or more short, singly labeled oligonucleotide probes. This makes each mRNA molecule visible as a computationally identifiable fluorescent spot by fluorescence microscopy. We demonstrate simultaneous detection of three mRNA species in single cells and mRNA detection in yeast, nematodes, fruit fly wing discs, and mammalian cell lines and neurons.

引用

收藏

页码：877 / 879

页数：3

相关论文

共 14 条

[1] Visualization of single RNA transcripts in situ [J].

Femino, A ;

Fay, FS ;

Fogarty, K ;

Singer, RH .

SCIENCE, 1998, 280 (5363) :585-590

[2]

Femino AM, 2003, METHOD ENZYMOL, V361, P245

[3] The GATA-factor elt-2 is essential for formation of the Caenorhabditis elegans intestine [J].

Fukushige, T ;

Hawkins, MG ;

McGhee, JD .

DEVELOPMENTAL BIOLOGY, 1998, 198 (02) :286-302

[4] DIFFERENTIAL SYNTHESIS OF GENES FOR RIBOSOMAL RNA DURING AMPHIBIAN OOGENESIS [J].

GALL, JG .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1968, 60 (02) :553-&

[5] Stochastic gene expression: from single molecules to the proteome [J].

Kaufmann, Benjamin B. ;

van Oudenaarden, Alexander .

CURRENT OPINION IN GENETICS & DEVELOPMENT, 2007, 17 (02) :107-112

[6] Fluorescence in situ hybridization: past, present and future [J].

Levsky, JM ;

Singer, RH .

JOURNAL OF CELL SCIENCE, 2003, 116 (14) :2833-2838

[7] Noise in gene expression determines cell fate in Bacillus subtilis [J].

Maamar, Hedia ;

Raj, Arjun ;

Dubnau, David .

SCIENCE, 2007, 317 (5837) :526-529

[8] ULTRA-SENSITIVE FISH USING PEROXIDASE-MEDIATED DEPOSITION OF BIOTIN-TYRAMIDE OR FLUOROCHROME-TYRAMIDE [J].

RAAP, AK ;

VANDECORPUT, MPC ;

VERVENNE, RAW ;

VANGIJLSWIJK, RPM ;

TANKE, HJ ;

WIEGANT, J .

HUMAN MOLECULAR GENETICS, 1995, 4 (04) :529-534

[9] Stochastic mRNA synthesis in mammalian cells [J].

Raj, Arjun ;

Peskin, Charles S. ;

Tranchina, Daniel ;

Vargas, Diana Y. ;

Tyagi, Sanjay .

PLOS BIOLOGY, 2006, 4 (10) :1707-1719

[10] Stability, specificity and fluorescence brightness of multiply-labeled fluorescent DNA probes [J].

Randolph, JB ;

Waggoner, AS .

NUCLEIC ACIDS RESEARCH, 1997, 25 (14) :2923-2929