Journal Article SC3-seq: A method for highly parallel and quantitative measurement of single-cell gene expression

Nakamura, Tomonori  ,  Yabuta, Yukihiro  ,  Okamoto, Ikuhiro  ,  Aramaki, Shinya  ,  Yokobayashi, Shihori  ,  Kurimoto, Kazuki  ,  Sekiguchi, Kiyotoshi  ,  Nakagawa, Masato  ,  Yamamoto, Takuya  ,  Saitou, Mitinori

43 ( 9 ) 2015-05-19 , Oxford University Press
Single-cell mRNA sequencing (RNA-seq) methods have undergone rapid development in recent years, and transcriptome analysis of relevant cell populations at single-cell resolution has become a key research area of biomedical sciences. We here present single-cell mRNA 3-prime end sequencing (SC3-seq), a practical methodology based on PCR amplification followed by 3-prime-end enrichment for highly quantitative, parallel and cost-effective measurement of gene expression in single cells. The SC3-seq allows excellent quantitative measurement of mRNAs ranging from the 10, 000-cell to 1-cell level, and accordingly, allows an accurate estimate of the transcript levels by a regression of the read counts of spike-in RNAs with defined copy numbers. The SC3-seq has clear advantages over other typical single-cell RNA-seq methodologies for the quantitative measurement of transcript levels and at a sequence depth required for the saturation of transcript detection. The SC3-seq distinguishes four distinct cell types in the peri-implantation mouse blastocysts. Furthermore, the SC3-seq reveals the heterogeneity in human-induced pluripotent stem cells (hiPSCs) cultured under on-feeder as well as feeder-free conditions, demonstrating a more homogeneous property of the feeder-free hiPSCs. We propose that SC3-seq might be used as a powerful strategy for single-cell transcriptome analysis in a broad range of investigations in biomedical sciences.

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