Reporter gene expression at single-cell level characterized with real-time RT-PCR, chemiluminescence, fluorescence, and electrochemical imaging

Hitoshi Shiku, Daisuke Okazaki, Junya Suzuki, Yasufumi Takahashi, Tatsuya Murata, Hidetaka Akita, Hideyoshi Harashima, Kosuke Ino, Tomokazu Matsue

Research output: Contribution to journalArticlepeer-review

8 Citations (Scopus)

Abstract

mRNA from single cells was quantified using real-time RT-PCR after recording the address and reporter protein activity with chemiluminescence, fluorescence, and electrochemical techniques, using luciferase, green fluorescent protein, and secreted alkaline phosphatase. mRNA copy number ranging from below 103 to 107 in single cells showed a lognormal distribution for both externally introduced reporter genes and internally expressed genes. The fluctuation in the gene expression decreased with the increase of the number of cells picked but did not decrease with the increase of mRNA copy number per cell. We found that the correlation coefficients for mRNA and protein expression in logarithmic plot at single-cell level were much lower than 1.00.

Original languageEnglish
Pages (from-to)4000-4008
Number of pages9
JournalFEBS Letters
Volume584
Issue number18
DOIs
Publication statusPublished - 2010 Sep

Keywords

  • Reporter assay
  • Single-cell analysis
  • Transcription
  • Translation

ASJC Scopus subject areas

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

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