TY - JOUR
T1 - Reporter gene expression at single-cell level characterized with real-time RT-PCR, chemiluminescence, fluorescence, and electrochemical imaging
AU - Shiku, Hitoshi
AU - Okazaki, Daisuke
AU - Suzuki, Junya
AU - Takahashi, Yasufumi
AU - Murata, Tatsuya
AU - Akita, Hidetaka
AU - Harashima, Hideyoshi
AU - Ino, Kosuke
AU - Matsue, Tomokazu
N1 - Funding Information:
This work was partly supported by a Grant-in-Aid for Scientific Research on Priority Areas (17066002) “Life Surveyor” from MEXT, of Japan; by a Grant-in-Aid for Scientific Research ( 18101006 , 21685008 , and 21106502 ) from MEXT ; and by a grant from the Center for Interdisciplinary Research, Tohoku University.
PY - 2010/9
Y1 - 2010/9
N2 - 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.
AB - 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.
KW - Reporter assay
KW - Single-cell analysis
KW - Transcription
KW - Translation
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U2 - 10.1016/j.febslet.2010.08.008
DO - 10.1016/j.febslet.2010.08.008
M3 - Article
C2 - 20699098
AN - SCOPUS:77956893951
VL - 584
SP - 4000
EP - 4008
JO - FEBS Letters
JF - FEBS Letters
SN - 0014-5793
IS - 18
ER -