In vivo monitoring of DNA vaccine gene expression using firefly luciferase as a naked DNA

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Jeon, Yong Hyun; Choi, Yun; Kang, Joo Hyun; Chung, June-Key; Lee, Yong Jin; Kim, Chul Woo; Jeong, Jae Min; Lee, Dong Soo; Lee, Myung Chul
Issue Date
Vaccine. 2006 Apr 12;24(16):3057-62. Epub 2006 Jan 30.
AnimalsFemale*Gene ExpressionGenes, ReporterImage Processing, Computer-AssistedLuciferases, Firefly/*analysis/geneticsLuminescent MeasurementsLymph Nodes/immunologyMiceMice, Inbred BALB CModels, AnimalRNA, Messenger/analysisReverse Transcriptase Polymerase Chain ReactionVaccines, DNA/administration & dosage/*genetics/*immunology
The administration of naked DNA into animals is increasing as a research tool to develop DNA vaccine. To monitor the distribution and duration of gene expression of a DNA vaccine in living organisms, we used the naked DNA encoding firefly luciferase (Fluc) as an imaging reporter gene, and evaluated in vivo bioluminescent images in a murine model. We observed bioluminescence at the injection site and at inguinal lymph node from 10h to 24h post-injection when DNA vaccine encoding Fluc (pcDNA3.1-Fluc) was injected into the bilateral posterior flanks in mice. Fluc gene expressions at injection sites and unilateral posterior flank inguinal lymph node were also confirmed by RT-PCR. However, when pcDNA3.1-Fluc was injected into the mid-dorsum bioluminescent signals were observed at the injection site for up to 14 days post-injection, but no bioluminescent signals were detected in inguinal lymph nodes. Concurrent mRNA expressions of Fluc gene at injection sites but not at inguinal lymph nodes were confirmed by RT-PCR. These findings suggest that optical imaging using Fluc could be useful for monitoring the location, intensity and duration of gene expression of naked DNA vaccines in living animals non-invasively and repetitively.
0264-410X (Print)
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College of Medicine/School of Medicine (의과대학/대학원)Nuclear Medicine (핵의학전공)Journal Papers (저널논문_핵의학전공)
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