Haemoglobin interferes with the ex vivo luciferase luminescence assay: consequence for detection of luciferase reporter gene expression in vivo
| Autor: | Charles Coutelle, Morvane Colin, S Moritz, M C Brahimi-Horn, Jacqueline Capeau, Holm Schneider |
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| Rok vydání: | 2000 |
| Předmět: |
Genetic Vectors
Gene Expression Biology Sensitivity and Specificity Adenoviridae Hemoglobins Mice In vivo Gene expression Genetics medicine Animals Luciferase Luciferases Lung Molecular Biology Reporter gene Genetic transfer Gene Transfer Techniques Transfection Molecular biology Perfusion Red blood cell medicine.anatomical_structure Liver Biochemistry Luminescent Measurements Molecular Medicine Spleen Ex vivo |
| Zdroj: | Gene Therapy. 7:1333-1336 |
| ISSN: | 1476-5462 0969-7128 |
| DOI: | 10.1038/sj.gt.3301248 |
| Popis: | The luciferase reporter gene is a useful tool for determining the efficacy of transfection of plasmid DNA and adenovirus-mediated gene transfer in vivo. However, we report here that the haemoglobin present in tissue samples can mask the detection of the luciferase activity and lead to underestimation of the luciferase levels. We evaluated the degree of interference in different organ samples of mice and investigated the possibilities for removal of haemoglobin from tissue samples by: (1) perfusion of the whole animal; (2) different hypotonic treatments lysing preferentially red blood cells; and (3) chromatographic separation. Removal of haemoglobin resulted in significantly improved detection of luciferase activity from tissue samples. The results indicate that the luciferase activity determined in tissue samples may not reflect the actual level of reporter gene expression, if contaminating blood is not taken into consideration. |
| Databáze: | OpenAIRE |
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