Journal article
Quantitative bioluminescence imaging of transgene expression in vivo
Analytical biochemistry, Vol.355(1), pp.90-94
2006
DOI: 10.1016/j.ab.2006.04.026
PMID: 16737677
Abstract
Bioluminescent imaging (BLI) is a widely used in vivo method to determine the location and relative intensity of luciferase expression in mice. Luciferase expression is observed following an i.p. dose of
d-luciferin, resulting in bioluminescence that is detected in anesthetized mice by a charge-coupled device camera. To establish whether BLI could be used as a quantitative measurement of non-viral-mediated luciferase expression, precise quantities of plasmid DNA encoding the luciferase gene were hydrodynamically dosed in mice. The results established a linear correlation between the DNA dose and the BLI response measured in liver which spanned five orders of magnitude. The level of luciferase expression was found to be a direct function of
d-luciferin dose. The time course of luciferase expression and the influence of multidosing of substrate were measured by BLI. The recovery of luciferase from the liver of hydrodynamically dosed mice allowed calibration of the BLI measurements. The results establish BLI’s limit-of-detection at 20
pg of luciferase per liver following a hydrodynamic dose of 100
pg of plasmid DNA. These results demonstrate that BLI is both sensitive and linear and should allow for the direct comparison of the efficiency of gene transfer vectors that target the liver.
Details
- Title: Subtitle
- Quantitative bioluminescence imaging of transgene expression in vivo
- Creators
- Garrett R RettigMarie McAnuffDijie LiuJi-Seon KimKevin G Rice
- Resource Type
- Journal article
- Publication Details
- Analytical biochemistry, Vol.355(1), pp.90-94
- DOI
- 10.1016/j.ab.2006.04.026
- PMID
- 16737677
- NLM abbreviation
- Anal Biochem
- ISSN
- 0003-2697
- eISSN
- 1096-0309
- Publisher
- Elsevier Inc
- Language
- English
- Date published
- 2006
- Academic Unit
- Radiology; Stead Family Department of Pediatrics; Pharmaceutical Sciences and Experimental Therapeutics; Craniofacial Anomalies Research Center; Medicinal and Natural Products Chemistry
- Record Identifier
- 9984065691102771
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