Quartz-Seq: a highly reproducible and sensitive single-cell RNA sequencing method, reveals non-genetic gene-expression heterogeneity

被引:336
作者
Sasagawa, Yohei [1 ,7 ]
Nikaido, Itoshi [1 ,7 ]
Hayashi, Tetsutaro [2 ]
Danno, Hiroki [3 ]
Uno, Kenichiro D. [1 ]
Imai, Takeshi [4 ,5 ]
Ueda, Hiroki R. [1 ,3 ,6 ]
机构
[1] RIKEN Ctr Dev Biol, Funct Genom Unit, Chuo Ku, Kobe, Hyogo 6500047, Japan
[2] RIKEN Ctr Dev Biol, Genome Resource & Anal Unit, Chuo Ku, Kobe, Hyogo 6500047, Japan
[3] RIKEN Ctr Dev Biol, Lab Syst Biol, Chuo Ku, Kobe, Hyogo 6500047, Japan
[4] RIKEN Ctr Dev Biol, Lab Sensory Circuit Format, Chuo Ku, Kobe, Hyogo 6500047, Japan
[5] JST, PRESTO, Chuo Ku, Kobe, Hyogo 6500047, Japan
[6] RIKEN, Quantitat Biol Ctr, Lab Synthet Biol, Chuo Ku, Kobe, Hyogo 6500047, Japan
[7] RIKEN, Adv Ctr Comp & Commun, Bioinformat Res Unit, Wako, Saitama 3510198, Japan
关键词
Single cell; RNA-seq; Transcriptome; Sequencing; Bioinformatics; Cellular heterogeneity; Cell biology; OLIGONUCLEOTIDE MICROARRAY ANALYSIS; ES CELLS; CDNA AMPLIFICATION; STEM-CELLS; NOISE; TRANSCRIPTOME; LANDSCAPE;
D O I
10.1186/gb-2013-14-4-r31
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Development of a highly reproducible and sensitive single-cell RNA sequencing (RNA-seq) method would facilitate the understanding of the biological roles and underlying mechanisms of non-genetic cellular heterogeneity. In this study, we report a novel single-cell RNA-seq method called Quartz-Seq that has a simpler protocol and higher reproducibility and sensitivity than existing methods. We show that single-cell Quartz-Seq can quantitatively detect various kinds of non-genetic cellular heterogeneity, and can detect different cell types and different cell-cycle phases of a single cell type. Moreover, this method can comprehensively reveal gene-expression heterogeneity between single cells of the same cell type in the same cell-cycle phase.
引用
收藏
页数:17
相关论文
共 30 条
[1]   Single-Cell Expression Analyses during Cellular Reprogramming Reveal an Early Stochastic and a Late Hierarchic Phase [J].
Buganim, Yosef ;
Faddah, Dina A. ;
Cheng, Albert W. ;
Itskovich, Elena ;
Markoulaki, Styliani ;
Ganz, Kibibi ;
Klemm, Sandy L. ;
van Oudenaarden, Alexander ;
Jaenisch, Rudolf .
CELL, 2012, 150 (06) :1209-1222
[2]   Single-Cell Analysis Reveals that Noncoding RNAs Contribute to Clonal Heterogeneity by Modulating Transcription Factor Recruitment [J].
Bumgarner, Stacie L. ;
Neuert, Gregor ;
Voight, Benjamin F. ;
Symbor-Nagrabska, Anna ;
Grisafi, Paula ;
van Oudenaarden, Alexander ;
Fink, Gerald R. .
MOLECULAR CELL, 2012, 45 (04) :470-482
[3]   An in situ hybridzation-based screen for heterogeneously expressed genes in mouse ES cells [J].
Carter, Mark G. ;
Stagg, Carole A. ;
Falco, Geppino ;
Yoshikawa, Toshiyuki ;
Bassey, Uwem C. ;
Aiba, Kazuhiro ;
Sharova, Lioudmila V. ;
Shaik, Nabeebi ;
Ko, Minoru S. H. .
GENE EXPRESSION PATTERNS, 2008, 8 (03) :181-198
[4]   Transcriptome-wide noise controls lineage choice in mammalian progenitor cells [J].
Chang, Hannah H. ;
Hemberg, Martin ;
Barahona, Mauricio ;
Ingber, Donald E. ;
Huang, Sui .
NATURE, 2008, 453 (7194) :544-U10
[5]   Regulatory activity revealed by dynamic correlations in gene expression noise [J].
Dunlop, Mary J. ;
Cox, Robert Sidney, III ;
Levine, Joseph H. ;
Murray, Richard M. ;
Elowitz, Michael B. .
NATURE GENETICS, 2008, 40 (12) :1493-1498
[6]   Functional roles for noise in genetic circuits [J].
Eldar, Avigdor ;
Elowitz, Michael B. .
NATURE, 2010, 467 (7312) :167-173
[7]   Highly Parallel Genome-Wide Expression Analysis of Single Mammalian Cells [J].
Fan, Jian-Bing ;
Chen, Jing ;
April, Craig S. ;
Fisher, Jeffrey S. ;
Klotzle, Brandy ;
Bibikova, Marina ;
Kaper, Fiona ;
Ronaghi, Mostafa ;
Linnarsson, Sten ;
Ota, Takayo ;
Chien, Jeremy ;
Laurent, Louise C. ;
Nisperos, Sean V. ;
Chen, Gina Y. ;
Zhong, Jiang F. .
PLOS ONE, 2012, 7 (02)
[8]   CEL-Seq: Single-Cell RNA-Seq by Multiplexed Linear Amplification [J].
Hashimshony, Tamar ;
Wagner, Florian ;
Sher, Noa ;
Yanai, Itai .
CELL REPORTS, 2012, 2 (03) :666-673
[9]  
Hausser J, 2009, J MACH LEARN RES, V10, P1469
[10]   Single-cell gene profiling of planarian stem cells using fluorescent activated cell sorting and its "index sorting" function for stem cell research [J].
Hayashi, Tetsutaro ;
Shibata, Norito ;
Okumura, Ryo ;
Kudome, Tomomi ;
Nishimura, Osamu ;
Tarui, Hiroshi ;
Agata, Kiyokazu .
DEVELOPMENT GROWTH & DIFFERENTIATION, 2010, 52 (01) :131-144