Haemoglobin interferes with the ex vivo luciferase luminescence assay: consequence for detection of luciferase reporter gene expression in vivo

被引：59
作者：
Colin, M
Moritz, S
Schneider, H
Capeau, J
Coutelle, C
Brahimi-Horn, MC
机构：
[1] Fac Med St Antoine, INSERM, U402, F-75571 Paris 12, France
[2] Univ London Imperial Coll Sci Technol & Med, Sch Med, Div Biomed Sci, Mol Genet Sect, London, England
来源：
GENE THERAPY | 2000年 / 7卷 / 15期
基金：
英国医学研究理事会;
关键词：
adenovirus; gene therapy; haemoglobin; luciferase; perfusion; red blood cells;
D O I：
10.1038/sj.gt.3301248
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
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.
引用
收藏
页码：1333 / 1336
页数：4
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