Abstract
Reporter genes produce a protein product in transfected cells that can be easily measured in intact or lysed cells and they have been extensively used in numerous basic and applied research applications. Over the past 10 years, reporter gene assays have been widely accepted and used for analysis of 2,3,7,8-tetrachlorodibenzo-p-dioxin and related dioxin-like compounds in various types of matrices, such as biological, environmental, food and feed samples, given that high-resolution instrumental analysis techniques are impractical for large-scale screening analysis. The most sensitive cell-based reporter gene bioassay systems developed are the mechanism-based CALUX (Chemically Activated Luciferase Expression) and CAFLUX (Chemically Activated Fluorescent Expression) bioassays, which utilize recombinant cell lines containing stably transfected dioxin (AhR)-responsive firefly luciferase or enhanced green fluorescent protein (EGFP) reporter genes, respectively. While the current CALUX and CAFLUX bioassays are very sensitive, increasing their lower limit of sensitivity, magnitude of response and dynamic range for chemical detection would significantly increase their utility, particularly for those samples that contain low levels of dioxin-like HAHs (i.e., serum). In this study, we report that the addition of modulators of cell signaling pathways or modification of cell culture conditions results in significant improvement in the magnitude and overall responsiveness of the existing CALUX and CAFLUX cell bioassays.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 1010-1016 |
| Number of pages | 7 |
| Journal | Science China Chemistry |
| Volume | 53 |
| Issue number | 5 |
| DOIs | |
| State | Published - May 2010 |
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Keywords
- Bioassay
- CAFLUX
- CALUX
- Dioxin
- Reporter gene
ASJC Scopus subject areas
- Chemistry(all)
Cite this
Enhancing the response of CALUX and CAFLUX cell bioassays for quantitative detection of dioxin-like compounds. / Zhao, Bin; Baston, David S.; Khan, Elaine; Sorrentino, Claudio; Denison, Michael S.
In: Science China Chemistry, Vol. 53, No. 5, 05.2010, p. 1010-1016.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Enhancing the response of CALUX and CAFLUX cell bioassays for quantitative detection of dioxin-like compounds
AU - Zhao, Bin
AU - Baston, David S.
AU - Khan, Elaine
AU - Sorrentino, Claudio
AU - Denison, Michael S.
PY - 2010/5
Y1 - 2010/5
N2 - Reporter genes produce a protein product in transfected cells that can be easily measured in intact or lysed cells and they have been extensively used in numerous basic and applied research applications. Over the past 10 years, reporter gene assays have been widely accepted and used for analysis of 2,3,7,8-tetrachlorodibenzo-p-dioxin and related dioxin-like compounds in various types of matrices, such as biological, environmental, food and feed samples, given that high-resolution instrumental analysis techniques are impractical for large-scale screening analysis. The most sensitive cell-based reporter gene bioassay systems developed are the mechanism-based CALUX (Chemically Activated Luciferase Expression) and CAFLUX (Chemically Activated Fluorescent Expression) bioassays, which utilize recombinant cell lines containing stably transfected dioxin (AhR)-responsive firefly luciferase or enhanced green fluorescent protein (EGFP) reporter genes, respectively. While the current CALUX and CAFLUX bioassays are very sensitive, increasing their lower limit of sensitivity, magnitude of response and dynamic range for chemical detection would significantly increase their utility, particularly for those samples that contain low levels of dioxin-like HAHs (i.e., serum). In this study, we report that the addition of modulators of cell signaling pathways or modification of cell culture conditions results in significant improvement in the magnitude and overall responsiveness of the existing CALUX and CAFLUX cell bioassays.
AB - Reporter genes produce a protein product in transfected cells that can be easily measured in intact or lysed cells and they have been extensively used in numerous basic and applied research applications. Over the past 10 years, reporter gene assays have been widely accepted and used for analysis of 2,3,7,8-tetrachlorodibenzo-p-dioxin and related dioxin-like compounds in various types of matrices, such as biological, environmental, food and feed samples, given that high-resolution instrumental analysis techniques are impractical for large-scale screening analysis. The most sensitive cell-based reporter gene bioassay systems developed are the mechanism-based CALUX (Chemically Activated Luciferase Expression) and CAFLUX (Chemically Activated Fluorescent Expression) bioassays, which utilize recombinant cell lines containing stably transfected dioxin (AhR)-responsive firefly luciferase or enhanced green fluorescent protein (EGFP) reporter genes, respectively. While the current CALUX and CAFLUX bioassays are very sensitive, increasing their lower limit of sensitivity, magnitude of response and dynamic range for chemical detection would significantly increase their utility, particularly for those samples that contain low levels of dioxin-like HAHs (i.e., serum). In this study, we report that the addition of modulators of cell signaling pathways or modification of cell culture conditions results in significant improvement in the magnitude and overall responsiveness of the existing CALUX and CAFLUX cell bioassays.
KW - Bioassay
KW - CAFLUX
KW - CALUX
KW - Dioxin
KW - Reporter gene
UR - http://www.scopus.com/inward/record.url?scp=77953329606&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=77953329606&partnerID=8YFLogxK
U2 - 10.1007/s11426-010-0142-8
DO - 10.1007/s11426-010-0142-8
M3 - Article
AN - SCOPUS:77953329606
VL - 53
SP - 1010
EP - 1016
JO - Science China Chemistry
JF - Science China Chemistry
SN - 1674-7291
IS - 5
ER -