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401. Quantitative Bioluminescence Imaging of Transgene Expression In Vivo
Abstract   Open access   Peer reviewed

401. Quantitative Bioluminescence Imaging of Transgene Expression In Vivo

Garrett Rettig, Marie Mcanuff, Dijie Liu, Ji-seon Kim and Kevin Rice
Molecular therapy, Vol.13(S1), p.S154
05/01/2006
DOI: 10.1016/j.ymthe.2006.08.463
url
https://doi.org/10.1016/j.ymthe.2006.08.463View
Published (Version of record) Open Access

Abstract

Bioluminescent imaging (BLI) is a widely used in vivo method to determine the location and relative intensity of luciferase expression in mice. In the present study, we have rigorously validated BLI to establish its utility as a quantitative measurement of luciferase expression for comparing the efficiency of non-viral gene transfer vectors in vivo. Precise quantities of plasmid DNA encoding the luciferase gene were hydrodynamically dosed in mice and luciferase expression was measured at different time intervals following an i.p. dose of D-luciferin. BLI provided a linear response when dosing 100 pg to 5 mg of plasmid DNA and integrating the light units measured in liver, resulting in a dynamic range that spanned 5- orders of magnitude. The level of luciferase expression was found to be a direct function of D-luciferin dose. BLI was used to serially sample mice to determine the time course of luciferase expression and to establish the influence of multi-dosing of substrate. BLI was found to be just as sensitive as a standard luciferase assay applied to homogenized mouse liver. The results establish a limit-of-detection of 20 pg of luciferase/liver following a hydrodynamic dose of 100 pg of plasmid DNA. BLI was also utilized to quantify the efficiency of knockdown of gene expression following siRNA dosing. The results demonstrate that BLI is both sensitive and linear and allows for the direct comparison of the efficiency of gene transfer vectors that target the liver.

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