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- Title
mRNA-Seq whole-transcriptome analysis of a single cell.
- Authors
Fuchou Tang; Barbacioru, Catalin; Yangzhou Wang; Nordman, Ellen; Clarence Lee; Nanlan Xu; Xiaohui Wang; Bodeau, John; Tuch, Brian B; Siddiqui, Asim; Lao, Kaiqin; Surani, M. Azim
- Abstract
Next-generation sequencing technology is a powerful tool for transcriptome analysis. However, under certain conditions, only a small amount of material is available, which requires more sensitive techniques that can preferably be used at the single-cell level. Here we describe a single-cell digital gene expression profiling assay. Using our mRNA-Seq assay with only a single mouse blastomere, we detected the expression of 75% (5,270) more genes than microarray techniques and identified 1,753 previously unknown splice junctions called by at least 5 reads. Moreover, 8–19% of the genes with multiple known transcript isoforms expressed at least two isoforms in the same blastomere or oocyte, which unambiguously demonstrated the complexity of the transcript variants at whole-genome scale in individual cells. Finally, for Dicer1−/− and Ago2−/− (Eif2c2−/−) oocytes, we found that 1,696 and 1,553 genes, respectively, were abnormally upregulated compared to wild-type controls, with 619 genes in common.
- Subjects
MESSENGER RNA; GENETIC regulation; GENE expression; GENOMES; BIOLOGICAL assay
- Publication
Nature Methods, 2009, Vol 6, Issue 5, p377
- ISSN
1548-7091
- Publication type
Article
- DOI
10.1038/nmeth.1315