KMID : 1148920090430050468
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Nuclear Medicine and Molecular Imaging 2009 Volume.43 No. 5 p.468 ~ p.477
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Imaging of Herpes Simplex Virus Type 1 Thymidine Kinase Gene Expression with Radiolabeled 5-(2-iodovinyl)-2¡¯-deoxyuridine (IVDU) in Liver by Hydrodynamic-based Procedure
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Song In-Ho
Lee Tae-Sup Kang Joo-Hyun Lee Yong-Jin Kim Kwang-Il An Gwang-Il Chung Wee-Sup Cheon Gi-Jeong Choi Chang-Woon Lim Sang-Moo
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Abstract
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Purpose: Hydrodynamic-based procedure is a simple and effective gene delivery method to lead a high gene expression in liver tissue. Non-invasive imaging reporter gene system has been used widely with herpes simplex virus type 1 thymidine kinase (HSV1-tk) and its various substrates. In the present study, we investigated to image the expression of HSV1-tk gene with 5-(2-iodovinyl)-2¡¯-deoxyuridine (IVDU) in mouse liver by the hydrodynamic- based procedure.
Materials and Methods: HSV1-tk or enhanced green fluorescence protein (EGFP) encoded plasmid DNA was transferred into the mouse liver by hydrodynaminc injection. At 24 h post-injection, RT-PCR, biodistribution, fluorescence imaging, nuclear imaging and digital wholebody autoradiography (DWBA) were performed to confirm transferred gene expression.
Results: In RT-PCR assay using mRNA from the mouse liver, specific bands of HSV1-tk and EGFP gene were observed in HSV1-tk and EGFP expressing plasmid injected mouse, respectively. Higher uptake of radiolabeled IVDU was exhibited in liver of HSV1-tk gene transferred mouse by biodistribution study. In fluorescence imaging, the liver showed specific fluorescence signal in EGFP gene transferred mouse. Gamma-camera image and DWBA results showed that radiolabeled IVDU was accumulated in the liver of HSV1-tk gene transferred mouse.
Conclusion: In this study, hydrodynamic-based procedure was effective in liver-specific gene delivery and it could be quantified with molecular imaging methods. Therefore, co-expression of HSV1-tk reporter gene and target gene by hydrodynamic-based procedure is expected to be a useful method for the evaluation of the target gene expression level with radiolabeled IVDU.
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KEYWORD
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Hydrodynamic-based procedure, HSV1-tk, IVDU, liver, molecular imaging
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