We are analyzing https://link.springer.com/article/10.1007/s00259-003-1441-5.

Title:
A novel triple-modality reporter gene for whole-body fluorescent, bioluminescent, and nuclear noninvasive imaging | European Journal of Nuclear Medicine and Molecular Imaging
Description:
Two genetic reporter systems were developed for multimodality reporter gene imaging of different molecular-genetic processes using fluorescence, bioluminescence (BLI), and nuclear imaging techniques. The eGFP cDNA was fused at the N-terminus with HSV1-tk cDNA bearing a nuclear export signal from MAPKK (NES-HSV1-tk) or with truncation at the N-terminus of the first 45 amino acids (Δ45HSV1-tk) and with firefly luciferase at the C-terminus. A single fusion protein with three functional subunits is formed following transcription and translation from a single open reading frame. The NES-TGL (NES-TGL) or Δ45HSV1-tk/GFP/luciferase (Δ45-TGL) triple-fusion gene cDNAs were cloned into a MoMLV-based retrovirus, which was used for transduction of U87 human glioma cells. The integrity, fluorescence, bioluminescence, and enzymatic activity of the TGL reporter proteins were assessed in vitro. The predicted molecular weight of the fusion proteins (~130 kDa) was confirmed by western blot. The U87-NES-TGL and U87-Δ45-TGL cells had cytoplasmic green fluorescence. The in vitro BLI was 7- and 13-fold higher in U87-NES-TGL and U87-Δ45-TGL cells compared to nontransduced control cells. The Ki of 14C-FIAU was 0.49±0.02, 0.51±0.03, and 0.003±0.001 ml/min/g in U87-NES-TGL, U87-Δ45-TGL, and wild-type U87 cells, respectively. Multimodality in vivo imaging studies were performed in nu/nu mice bearing multiple s.c. xenografts established from U87-NES-TGL, U87-Δ45-TGL, and wild-type U87 cells. BLI was performed after administration of d-luciferin (150 mg/kg i.v.). Gamma camera or PET imaging was conducted at 2 h after i.v. administration of [131I]FIAU (7.4 MBq/animal) or [124I]FIAU (7.4 MBq/animal), respectively. Whole-body fluorescence imaging was performed in parallel with the BLI and radiotracer imaging studies. In vivo BLI and gamma camera imaging showed specific localization of luminescence and radioactivity to the TGL transduced xenografts with background levels of activity in the wild-type xenografts. Tissue sampling yielded values of 0.47%±0.08%, 0.86%±0.06%, and 0.03%±0.01%dose/g [131I]FIAU in U87-NES-TGL, U87-Δ45-TGL, and U87 xenografts, respectively. The TGL triple-fusion reporter gene preserves the functional activity of its subunits and is very effective for multimodality imaging. It provides for the seamless transition from fluorescence microscopy and FACS to whole-body bioluminescence imaging, to nuclear (PET, SPET, gamma camera) imaging, and back to in situ fluorescence image analysis.
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Keywords {🔍}

imaging, google, scholar, pubmed, cas, gene, article, tjuvajev, reporter, blasberg, expression, balatoni, ponomarev, thymidine, cancer, doubrovin, gelovani, virus, kinase, finn, noninvasive, cells, vivo, mice, fluorescent, beresten, green, herpes, gambhir, nuclear, serganova, simplex, med, res, molecular, protein, cell, larson, sadelain, positron, emission, tomography, ageyeva, fluorescence, human, access, nucl, living, mol, fusion,

Topics {✒️}

monitoring e-selectin-mediated adhesion month download article/chapter hsv-1 thymidine kinase u87-δ45-tgl cells compared herpes virus infection wild-type u87 cells triple-fusion gene cdnas renilla luciferase-aequorea gfp hsv1-tk cdna bearing triple-modality reporter gene δ45hsv1-tk/gfp/luciferase juri gelovani tjuvajev truncated hsv1-tk green fluorescent protein intact blood-brain barrier invasive molecular-genetic imaging protein-protein interactions u87-δ45-tgl cells single fusion protein combined pet/ct scanner red fluorescent proteins fusion reporter gene vesicular stomatitis virus article european journal wild-type xenografts full article pdf reporter gene expression genetic reporter systems nes-hsv1-tk p53-dependent genes reporter gene imaging ubiquitous green cells transgene expression mediated molecular-genetic targets author correspondence multiple reporter genes marker/reporter gene immunogenic reporter gene tgl reporter proteins privacy choices/manage cookies gelovani tjuvajev jg gene transfer strategies gene transfer studies stable gene transfer related subjects human tumor cells endogenous gene expression molecular-genetic processes tgl transduced xenografts fusion protein

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         headline:A novel triple-modality reporter gene for whole-body fluorescent, bioluminescent, and nuclear noninvasive imaging
         description:Two genetic reporter systems were developed for multimodality reporter gene imaging of different molecular-genetic processes using fluorescence, bioluminescence (BLI), and nuclear imaging techniques. The eGFP cDNA was fused at the N-terminus with HSV1-tk cDNA bearing a nuclear export signal from MAPKK (NES-HSV1-tk) or with truncation at the N-terminus of the first 45 amino acids (Δ45HSV1-tk) and with firefly luciferase at the C-terminus. A single fusion protein with three functional subunits is formed following transcription and translation from a single open reading frame. The NES-TGL (NES-TGL) or Δ45HSV1-tk/GFP/luciferase (Δ45-TGL) triple-fusion gene cDNAs were cloned into a MoMLV-based retrovirus, which was used for transduction of U87 human glioma cells. The integrity, fluorescence, bioluminescence, and enzymatic activity of the TGL reporter proteins were assessed in vitro. The predicted molecular weight of the fusion proteins (~130 kDa) was confirmed by western blot. The U87-NES-TGL and U87-Δ45-TGL cells had cytoplasmic green fluorescence. The in vitro BLI was 7- and 13-fold higher in U87-NES-TGL and U87-Δ45-TGL cells compared to nontransduced control cells. The Ki of 14C-FIAU was 0.49±0.02, 0.51±0.03, and 0.003±0.001 ml/min/g in U87-NES-TGL, U87-Δ45-TGL, and wild-type U87 cells, respectively. Multimodality in vivo imaging studies were performed in nu/nu mice bearing multiple s.c. xenografts established from U87-NES-TGL, U87-Δ45-TGL, and wild-type U87 cells. BLI was performed after administration of d-luciferin (150 mg/kg i.v.). Gamma camera or PET imaging was conducted at 2 h after i.v. administration of [131I]FIAU (7.4 MBq/animal) or [124I]FIAU (7.4 MBq/animal), respectively. Whole-body fluorescence imaging was performed in parallel with the BLI and radiotracer imaging studies. In vivo BLI and gamma camera imaging showed specific localization of luminescence and radioactivity to the TGL transduced xenografts with background levels of activity in the wild-type xenografts. Tissue sampling yielded values of 0.47%±0.08%, 0.86%±0.06%, and 0.03%±0.01%dose/g [131I]FIAU in U87-NES-TGL, U87-Δ45-TGL, and U87 xenografts, respectively. The TGL triple-fusion reporter gene preserves the functional activity of its subunits and is very effective for multimodality imaging. It provides for the seamless transition from fluorescence microscopy and FACS to whole-body bioluminescence imaging, to nuclear (PET, SPET, gamma camera) imaging, and back to in situ fluorescence image analysis.
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            Herpes virus type one
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            Nuclear Medicine
            Imaging / Radiology
            Orthopedics
            Cardiology
            Oncology
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      headline:A novel triple-modality reporter gene for whole-body fluorescent, bioluminescent, and nuclear noninvasive imaging
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         Multimodality imaging
         Herpes virus type one
         Thymidine kinase
         Green fluorescent protein
         Luciferase
         FIAU
         Nuclear Medicine
         Imaging / Radiology
         Orthopedics
         Cardiology
         Oncology
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