MRNA-Seq Whole-transcriptome Analysis of a Single Cell

Overview

Journal Nat Methods

Specialties Biomedical Engineering
Pathology

Date 2009 Apr 8

PMID 19349980

Citations 1640

Authors

Fuchou Tang

Catalin Barbacioru

Yangzhou Wang

Ellen Nordman

Clarence Lee

Nanlan Xu

Xiaohui Wang

John Bodeau

Brian B Tuch

Asim Siddiqui

Kaiqin Lao

M Azim Surani

Affiliations

Soon will be listed here.

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.

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