研究人员在日前在线出版的《自然—方法学》期刊报告说，下一代的测序仪将能够读出单个细胞的全部核糖核酸分子池（RNA pool），即转录组。

转录组提供了有关活性基因的功能、调控和互相依赖的信息。最近，科学家们将高通量测序仪应用于转录组分析（RNA-Seq），新方法让研究人员能够更为详细地了解有多少基因组被转录、每个基因产生了多少RNA变异。这种转录组分析方法的局限是它需要成百上千的细胞。

Azim Surani和同事将转录组分析方法应用到单个细胞的分析中，并以前所未有的精度分析了取自四细胞胚胎和卵母细胞的单个老鼠细胞的转录组。

新方法不仅对发育生物学的研究至关重要，而且也让科学家们能够在胚胎发育的早期深入研究单个细胞中全部基因表达、研究复杂组织中的细胞异质性，并在单个细胞的水平上分析疾病的发展。（生物谷Bioon.com）

生物谷推荐原始出处：

Nature Methods 6, 377 - 382 (2009) 19 April 2009 | doi:10.1038/nmeth.1315

mRNA-Seq whole-transcriptome analysis of a single cell

Fuchou Tang1,3, Catalin Barbacioru2,3, Yangzhou Wang2, Ellen Nordman2, Clarence Lee2, Nanlan Xu2, Xiaohui Wang2, John Bodeau2, Brian B Tuch2, Asim Siddiqui2, Kaiqin Lao2 & M Azim Surani1

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.

1 Wellcome Trust–Cancer Research UK Gurdon Institute of Cancer and Developmental Biology, University of Cambridge, Cambridge, UK.
2 Molecular Cell Biology Division, Applied Biosystems, Foster City, California, USA.
3 These authors contributed equally to this work.