Highly parallel genome-wide expression analysis of single mammalian cells.

PloS One
Jian-Bing FanJiang F Zhong

Abstract

We have developed a high-throughput amplification method for generating robust gene expression profiles using single cell or low RNA inputs. The method uses tagged priming and template-switching, resulting in the incorporation of universal PCR priming sites at both ends of the synthesized cDNA for global PCR amplification. Coupled with a whole-genome gene expression microarray platform, we routinely obtain expression correlation values of R(2)~0.76-0.80 between individual cells and R(2)~0.69 between 50 pg total RNA replicates. Expression profiles generated from single cells or 50 pg total RNA correlate well with that generated with higher input (1 ng total RNA) (R(2)~0.80). Also, the assay is sufficiently sensitive to detect, in a single cell, approximately 63% of the number of genes detected with 1 ng input, with approximately 97% of the genes detected in the single-cell input also detected in the higher input. In summary, our method facilitates whole-genome gene expression profiling in contexts where starting material is extremely limiting, particularly in areas such as the study of progenitor cells in early development and tumor stem cell biology.

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Citations

Feb 6, 2014·The Analyst·Kerstin GallerUte Neugebauer
Nov 26, 2015·Fertility and Sterility·Eleni MantikouMark de Jong
Dec 15, 2015·Scientific Reports·Yiqiang ZhangCharles Wang
Jan 1, 2014·Journal of Clinical Medicine·Eugene Pergament
Oct 27, 2017·Oncotarget·Andres StuckyJiang F Zhong
May 3, 2018·Carcinogenesis·Xuelian ChenJiang F Zhong

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Datasets Mentioned

BETA
GSE34365

Methods Mentioned

BETA
PCR
Assay

Software Mentioned

DAVID
GenomeStudio®

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