Transcriptome analysis by strand-specific sequencing of complementary DNA

被引:676
作者
Parkhomchuk, Dmitri [1 ]
Borodina, Tatiana [1 ]
Amstislavskiy, Vyacheslav [1 ]
Banaru, Maria [1 ]
Hallen, Linda [1 ]
Krobitsch, Sylvia [1 ]
Lehrach, Hans [1 ]
Soldatov, Alexey [1 ]
机构
[1] Max Planck Inst Mol Genet, Dept Vertebrate Genom, D-14195 Berlin, Germany
关键词
RNA; MAPS;
D O I
10.1093/nar/gkp596
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
High-throughput complementary DNA sequencing (RNA-Seq) is a powerful tool for whole-transcriptome analysis, supplying information about a transcript's expression level and structure. However, it is difficult to determine the polarity of transcripts, and therefore identify which strand is transcribed. Here, we present a simple cDNA sequencing protocol that preserves information about a transcript's direction. Using Saccharomyces cerevisiae and mouse brain transcriptomes as models, we demonstrate that knowing the transcript's orientation allows more accurate determination of the structure and expression of genes. It also helps to identify new genes and enables studying promoter-associated and antisense transcription. The transcriptional landscapes we obtained are available online.
引用
收藏
页数:7
相关论文
共 21 条
[1]   Identification and analysis of functional elements in 1% of the human genome by the ENCODE pilot project [J].
Birney, Ewan ;
Stamatoyannopoulos, John A. ;
Dutta, Anindya ;
Guigo, Roderic ;
Gingeras, Thomas R. ;
Margulies, Elliott H. ;
Weng, Zhiping ;
Snyder, Michael ;
Dermitzakis, Emmanouil T. ;
Stamatoyannopoulos, John A. ;
Thurman, Robert E. ;
Kuehn, Michael S. ;
Taylor, Christopher M. ;
Neph, Shane ;
Koch, Christoph M. ;
Asthana, Saurabh ;
Malhotra, Ankit ;
Adzhubei, Ivan ;
Greenbaum, Jason A. ;
Andrews, Robert M. ;
Flicek, Paul ;
Boyle, Patrick J. ;
Cao, Hua ;
Carter, Nigel P. ;
Clelland, Gayle K. ;
Davis, Sean ;
Day, Nathan ;
Dhami, Pawandeep ;
Dillon, Shane C. ;
Dorschner, Michael O. ;
Fiegler, Heike ;
Giresi, Paul G. ;
Goldy, Jeff ;
Hawrylycz, Michael ;
Haydock, Andrew ;
Humbert, Richard ;
James, Keith D. ;
Johnson, Brett E. ;
Johnson, Ericka M. ;
Frum, Tristan T. ;
Rosenzweig, Elizabeth R. ;
Karnani, Neerja ;
Lee, Kirsten ;
Lefebvre, Gregory C. ;
Navas, Patrick A. ;
Neri, Fidencio ;
Parker, Stephen C. J. ;
Sabo, Peter J. ;
Sandstrom, Richard ;
Shafer, Anthony .
NATURE, 2007, 447 (7146) :799-816
[2]   Stem cell transcriptome profiling via massive-scale mRNA sequencing [J].
Cloonan, Nicole ;
Forrest, Alistair R. R. ;
Kolle, Gabriel ;
Gardiner, Brooke B. A. ;
Faulkner, Geoffrey J. ;
Brown, Mellissa K. ;
Taylor, Darrin F. ;
Steptoe, Anita L. ;
Wani, Shivangi ;
Bethel, Graeme ;
Robertson, Alan J. ;
Perkins, Andrew C. ;
Bruce, Stephen J. ;
Lee, Clarence C. ;
Ranade, Swati S. ;
Peckham, Heather E. ;
Manning, Jonathan M. ;
McKernan, Kevin J. ;
Grimmond, Sean M. .
NATURE METHODS, 2008, 5 (07) :613-619
[3]   Nascent RNA Sequencing Reveals Widespread Pausing and Divergent Initiation at Human Promoters [J].
Core, Leighton J. ;
Waterfall, Joshua J. ;
Lis, John T. .
SCIENCE, 2008, 322 (5909) :1845-1848
[4]   Highly expressed genes are associated with inverse antisense transcription in mouse [J].
Gyoerffy, Andras ;
Surowiak, Pawel ;
Tulassay, Zsolt ;
Gyoerffy, Balazs .
JOURNAL OF GENETICS, 2007, 86 (02) :103-109
[5]   The Antisense Transcriptomes of Human Cells [J].
He, Yiping ;
Vogelstein, Bert ;
Velculescu, Victor E. ;
Papadopoulos, Nickolas ;
Kinzler, Kenneth W. .
SCIENCE, 2008, 322 (5909) :1855-1857
[6]   Archaeal dUTPase enhances PCR amplifications with archaeal DNA polymerases by preventing dUTP incorporation [J].
Hogrefe, HH ;
Hansen, CJ ;
Scott, BR ;
Nielson, KB .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2002, 99 (02) :596-601
[7]   Genome-wide transcription and the implications for genomic organization [J].
Kapranov, Philipp ;
Willingham, Aarron T. ;
Gingeras, Thomas R. .
NATURE REVIEWS GENETICS, 2007, 8 (06) :413-423
[8]   RNA maps reveal new RNA classes and a possible function for pervasive transcription [J].
Kapranov, Philipp ;
Cheng, Jill ;
Dike, Sujit ;
Nix, David A. ;
Duttagupta, Radharani ;
Willingham, Aarron T. ;
Stadler, Peter F. ;
Hertel, Jana ;
Hackermueller, Joerg ;
Hofacker, Ivo L. ;
Bell, Ian ;
Cheung, Evelyn ;
Drenkow, Jorg ;
Dumais, Erica ;
Patel, Sandeep ;
Helt, Gregg ;
Ganesh, Madhavan ;
Ghosh, Srinka ;
Piccolboni, Antonio ;
Sementchenko, Victor ;
Tammana, Hari ;
Gingeras, Thomas R. .
SCIENCE, 2007, 316 (5830) :1484-1488
[9]   Highly integrated single-base resolution maps of the epigenome in Arabidopsis [J].
Lister, Ryan ;
O'Malley, Ronan C. ;
Tonti-Filippini, Julian ;
Gregory, Brian D. ;
Berry, Charles C. ;
Millar, A. Harvey ;
Ecker, Joseph R. .
CELL, 2008, 133 (03) :523-536
[10]   RNA-seq: An assessment of technical reproducibility and comparison with gene expression arrays [J].
Marioni, John C. ;
Mason, Christopher E. ;
Mane, Shrikant M. ;
Stephens, Matthew ;
Gilad, Yoav .
GENOME RESEARCH, 2008, 18 (09) :1509-1517