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Direct RNA sequencing.

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Title: Direct RNA sequencing.
Authors: Ozsolak, Fatih, Platt, Adam R., Jones, Dan R., Reifenberger, Jeffrey G., Sass, Lauryn E., McInerney, Peter, Thompson, John F., Bowers, Jayson, Jarosz, Mirna, Milos, Patrice M.
Source: Nature. 10/8/2009, Vol. 461 Issue 7265, p814-818. 5p. 4 Graphs.
Subjects: Human physiology, Diseases, Genomes, Protein microarrays, RNA, Antisense DNA, Saccharomyces cerevisiae, Oligonucleotides, Heterogeneity
Abstract: Our understanding of human biology and disease is ultimately dependent on a complete understanding of the genome and its functions. The recent application of microarray and sequencing technologies to transcriptomics has changed the simplistic view of transcriptomes to a more complicated view of genome-wide transcription where a large fraction of transcripts emanates from unannotated parts of genomes, and underlined our limited knowledge of the dynamic state of transcription. Most of this broad body of knowledge was obtained indirectly because current transcriptome analysis methods typically require RNA to be converted to complementary DNA (cDNA) before measurements, even though the cDNA synthesis step introduces multiple biases and artefacts that interfere with both the proper characterization and quantification of transcripts. Furthermore, cDNA synthesis is not particularly suitable for the analysis of short, degraded and/or small quantity RNA samples. Here we report direct single molecule RNA sequencing without prior conversion of RNA to cDNA. We applied this technology to sequence femtomole quantities of poly(A)+ Saccharomyces cerevisiae RNA using a surface coated with poly(dT) oligonucleotides to capture the RNAs at their natural poly(A) tails and initiate sequencing by synthesis. We observed transcript 3′ end heterogeneity and polyadenylated small nucleolar RNAs. This study provides a path to high-throughput and low-cost direct RNA sequencing and achieving the ultimate goal of a comprehensive and bias-free understanding of transcriptomes. [ABSTRACT FROM AUTHOR]
Copyright of Nature is the property of Springer Nature and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
Database: Psychology and Behavioral Sciences Collection
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  Data: <searchLink fieldCode="AR" term="%22Ozsolak%2C+Fatih%22">Ozsolak, Fatih</searchLink><br /><searchLink fieldCode="AR" term="%22Platt%2C+Adam+R%2E%22">Platt, Adam R.</searchLink><br /><searchLink fieldCode="AR" term="%22Jones%2C+Dan+R%2E%22">Jones, Dan R.</searchLink><br /><searchLink fieldCode="AR" term="%22Reifenberger%2C+Jeffrey+G%2E%22">Reifenberger, Jeffrey G.</searchLink><br /><searchLink fieldCode="AR" term="%22Sass%2C+Lauryn+E%2E%22">Sass, Lauryn E.</searchLink><br /><searchLink fieldCode="AR" term="%22McInerney%2C+Peter%22">McInerney, Peter</searchLink><br /><searchLink fieldCode="AR" term="%22Thompson%2C+John+F%2E%22">Thompson, John F.</searchLink><br /><searchLink fieldCode="AR" term="%22Bowers%2C+Jayson%22">Bowers, Jayson</searchLink><br /><searchLink fieldCode="AR" term="%22Jarosz%2C+Mirna%22">Jarosz, Mirna</searchLink><br /><searchLink fieldCode="AR" term="%22Milos%2C+Patrice+M%2E%22">Milos, Patrice M.</searchLink>
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  Data: Our understanding of human biology and disease is ultimately dependent on a complete understanding of the genome and its functions. The recent application of microarray and sequencing technologies to transcriptomics has changed the simplistic view of transcriptomes to a more complicated view of genome-wide transcription where a large fraction of transcripts emanates from unannotated parts of genomes, and underlined our limited knowledge of the dynamic state of transcription. Most of this broad body of knowledge was obtained indirectly because current transcriptome analysis methods typically require RNA to be converted to complementary DNA (cDNA) before measurements, even though the cDNA synthesis step introduces multiple biases and artefacts that interfere with both the proper characterization and quantification of transcripts. Furthermore, cDNA synthesis is not particularly suitable for the analysis of short, degraded and/or small quantity RNA samples. Here we report direct single molecule RNA sequencing without prior conversion of RNA to cDNA. We applied this technology to sequence femtomole quantities of poly(A)+ Saccharomyces cerevisiae RNA using a surface coated with poly(dT) oligonucleotides to capture the RNAs at their natural poly(A) tails and initiate sequencing by synthesis. We observed transcript 3′ end heterogeneity and polyadenylated small nucleolar RNAs. This study provides a path to high-throughput and low-cost direct RNA sequencing and achieving the ultimate goal of a comprehensive and bias-free understanding of transcriptomes. [ABSTRACT FROM AUTHOR]
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  Data: <i>Copyright of Nature is the property of Springer Nature and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.)
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              Text: 10/8/2009
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