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Proc Natl Acad Sci U S A. 2008 Dec 23;105(51):20173-8. doi: 10.1073/pnas.0806355106. Epub 2008 Dec 15.

Integrated microfluidic bioprocessor for single-cell gene expression analysis.

Author information

1
University of California San Francisco/University of California Berkeley Joint Graduate Group in Bioengineering, Lawrence Berkeley National Laboratory, University of California, Berkeley, CA 94720, USA.

Abstract

An integrated microdevice is developed for the analysis of gene expression in single cells. The system captures a single cell, transcribes and amplifies the mRNA, and quantitatively analyzes the products of interest. The key components of the microdevice include integrated nanoliter metering pumps, a 200-nL RT-PCR reactor with a single-cell capture pad, and an affinity capture matrix for the purification and concentration of products that is coupled to a microfabricated capillary electrophoresis separation channel for product analysis. Efficient microchip integration of these processes enables the sensitive and quantitative examination of gene expression variation at the single-cell level. This microdevice is used to measure siRNA knockdown of the GAPDH gene in individual Jurkat cells. Single-cell measurements suggests the presence of 2 distinct populations of cells with moderate (approximately 50%) or complete (approximately 0%) silencing. This stochastic variation in gene expression and silencing within single cells is masked by conventional bulk measurements.

PMID:
19075237
PMCID:
PMC2629289
DOI:
10.1073/pnas.0806355106
[Indexed for MEDLINE]
Free PMC Article

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