RT Journal Article
SR Electronic
T1 Production of transgenic mice expressing firefly luciferase gene controlled by α-fetoprotein enhancer/promoter
JF Journal of Nuclear Medicine
JO J Nucl Med
FD Society of Nuclear Medicine
SP 1020
OP 1020
VO 50
IS supplement 2
A1 Park, JH
A1 Kang, JH
A1 Lee, TS
A1 Kim, KI
A1 Park, YS
A1 Choi, CW
A1 Lim, SM
YR 2009
UL http://jnm.snmjournals.org/content/50/supplement_2/1020.abstract
AB 1020 Objectives Development of a small animal imaging system for hepatocellular carcinoma(HCC) specific reporter gene expression will enable us to image carcinogenesis or monitor therapeutic intervention of HCC in vivo. Fetoprotein(AFP) is a fetal protein which is not expressed after birth but is re-expressed in HCC. In this study, we developed a firefly luciferase(fLuc) transgenic mouse in which fLuc expression is controlled by AFP enhancer/promoter only in AFP producing cells. Methods pGL3-AFPfLuc containing fLuc gene driven by AFP enhancer/promoter was constructed. For the verification of this system pGL3-AFPfLuc was transiently transfected to AFP-producing HCC cells(HuH-7) and AFP-non producing cells(C6) using lipofectamine. The expression of fLuc gene was measured by fLuc activity assay. pGL3-AFPfLuc was used for the production of fLuc-transgenic mice. The propriety of the transgenic mouse model was evaluated by in vivo bioluminescent imaging (BLI) in fetus or neonatal mice. Results After transfection of pGL3-AFPfLuc, the fLuc activity was observed in AFP-producing HCC cells, but not in AFP-nonproducing cells. Eleven positive founders among 110 founders were obtained using genomic PCR typing and 8 of 11 lines were male. Eight male genotype positive founders were mated with wild type female mice and its progenies of 3 lines(39, 59 and 78) showed strong bioluminescence signal in their liver region at 1 day after birth as well as fetus. Normal fetus and newborn mice did not express fLuc in their liver region. Conclusions We developed a transgenic mouse model to image AFP-producing cells with BLI using AFP enhancer/promoter and fLuc.