BMC Genomics | |
The complex transcriptional landscape of the anucleate human platelet | |
Research Article | |
Joan Kupper1  Kathleen Delgrosso1  Adam Ertel1  Paolo Fortina1  Leonard C Edelstein2  Paul F Bray2  Steven E McKenzie2  Srikanth Nagalla2  Isidore Rigoutsos3  Eric Londin3  Huang-Wen Chen3  Phillipe Loher3  Yi Jing3  | |
[1] Cancer Genomics Laboratory, Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, PA, USA;Cardeza Foundation for Hematologic Research, Division of Hematology, Department of Medicine, Thomas Jefferson University, Philadelphia, PA, USA;Computational Medicine Center, Thomas Jefferson University, Philadelphia, PA, USA; | |
关键词: Platelet; Transcriptome; Ribosomal RNA; Non-coding RNA; miRNA; Repeat elements; Antisense transcripts; | |
DOI : 10.1186/1471-2164-14-1 | |
received in 2012-06-28, accepted in 2012-12-05, 发布年份 2013 | |
来源: Springer | |
【 摘 要 】
BackgroundHuman blood platelets are essential to maintaining normal hemostasis, and platelet dysfunction often causes bleeding or thrombosis. Estimates of genome-wide platelet RNA expression using microarrays have provided insights to the platelet transcriptome but were limited by the number of known transcripts. The goal of this effort was to deep-sequence RNA from leukocyte-depleted platelets to capture the complex profile of all expressed transcripts.ResultsFrom each of four healthy individuals we generated long RNA (≥40 nucleotides) profiles from total and ribosomal-RNA depleted RNA preparations, as well as short RNA (<40 nucleotides) profiles. Analysis of ~1 billion reads revealed that coding and non-coding platelet transcripts span a very wide dynamic range (≥16 PCR cycles beyond β-actin), a result we validated through qRT-PCR on many dozens of platelet messenger RNAs. Surprisingly, ribosomal-RNA depletion significantly and adversely affected estimates of the relative abundance of transcripts. Of the known protein-coding loci, ~9,500 are present in human platelets. We observed a strong correlation between mRNAs identified by RNA-seq and microarray for well-expressed mRNAs, but RNASeq identified many more transcripts of lower abundance and permitted discovery of novel transcripts.ConclusionsOur analyses revealed diverse classes of non-coding RNAs, including: pervasive antisense transcripts to protein-coding loci; numerous, previously unreported and abundant microRNAs; retrotransposons; and thousands of novel un-annotated long and short intronic transcripts, an intriguing finding considering the anucleate nature of platelets. The data are available through a local mirror of the UCSC genome browser and can be accessed at:http://cm.jefferson.edu/platelets_2012/.
【 授权许可】
CC BY
© Bray et al.; licensee BioMed Central Ltd. 2013
【 预 览 】
Files | Size | Format | View |
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RO202311091504050ZK.pdf | 940KB | download |
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