Quantitative single-cell gene expression measurements of multiple genes in response to hypoxia treatment

Jia Zeng, Jiangxin Wang, Weimin Gao, Aida Mohammadreza, Laimonas Kelbauskas, Weiwen Zhang, Roger H. Johnson, Deirdre Meldrum

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Cell-to-cell heterogeneity in gene transcription plays a central role in a variety of vital cell processes. To quantify gene expression heterogeneity patterns among cells and to determine their biological significance, methods to measure gene expression levels at the single-cell level are highly needed. We report an experimental technique based on the DNA-intercalating fluorescent dye SYBR green for quantitative expression level analysis of up to ten selected genes in single mammalian cells. The method features a two-step procedure consisting of a step to isolate RNA from a single mammalian cell, synthesize cDNA from it, and a qPCR step. We applied the method to cell populations exposed to hypoxia, quantifying expression levels of seven different genes spanning a wide dynamic range of expression in randomly picked single cells. In the experiment, 72 single Barrett's esophageal epithelial (CP-A) cells, 36 grown under normal physiological conditions (controls) and 36 exposed to hypoxia for 30 min, were randomly collected and used for measuring the expression levels of 28S rRNA, PRKAA1, GAPDH, Angptl4, MT3, PTGES, and VEGFA genes. The results demonstrate that the method is sensitive enough to measure alterations in gene expression at the single-cell level, clearly showing heterogeneity within a cell population. We present technical details of the method development and implementation, and experimental results obtained by use of the procedure. We expect the advantages of this technique will facilitate further developments and advances in the field of single-cell gene expression profiling on a nanotechnological scale, and eventually as a tool for future point-of-care medical applications.

Original languageEnglish (US)
Pages (from-to)3-13
Number of pages11
JournalAnalytical and Bioanalytical Chemistry
Volume401
Issue number1
DOIs
StatePublished - Jul 2011

Fingerprint

Gene expression
Genes
Cells
Gene Expression
Medical applications
Transcription
Fluorescent Dyes
Complementary DNA
RNA
Hypoxia
DNA
Point-of-Care Systems
Experiments
Gene Expression Profiling
Population

Keywords

  • Gene expression
  • Hypoxia treatment
  • Single-cell

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry

Cite this

Quantitative single-cell gene expression measurements of multiple genes in response to hypoxia treatment. / Zeng, Jia; Wang, Jiangxin; Gao, Weimin; Mohammadreza, Aida; Kelbauskas, Laimonas; Zhang, Weiwen; Johnson, Roger H.; Meldrum, Deirdre.

In: Analytical and Bioanalytical Chemistry, Vol. 401, No. 1, 07.2011, p. 3-13.

Research output: Contribution to journalArticle

Zeng, Jia ; Wang, Jiangxin ; Gao, Weimin ; Mohammadreza, Aida ; Kelbauskas, Laimonas ; Zhang, Weiwen ; Johnson, Roger H. ; Meldrum, Deirdre. / Quantitative single-cell gene expression measurements of multiple genes in response to hypoxia treatment. In: Analytical and Bioanalytical Chemistry. 2011 ; Vol. 401, No. 1. pp. 3-13.
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