mRNA-Seq whole-transcriptome analysis of a single cell

作者：Tang Fuchou; Barbacioru Catalin; Wang Yangzhou; Nordman Ellen; Lee Clarence; Xu Nanlan; Wang Xiaohui; Bodeau John; Tuch Brian B; Siddiqui Asim; Lao Kaiqin^*; Surani M Azim

来源：Nature Methods, 2009, 6(5): 377-U86.

DOI：10.1038/NMETH.1315

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.

• 出版日期2009-5