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Demonstration of Tightly Radiation-Controlled Molecular Switch Based on CArG Repeats by In Vivo Molecular Imaging
Hsieh Y.-J.2; Hwu L.4; Ke C.-C.1; Huang A.-L.1; Chen F.-D.1; Wu S.-J.2; Chen S.C.-J.2; Zhao Y.-H.7; Liu R.-S.4
2015-12-01
Source PublicationMolecular Imaging and Biology
ISSN18602002 15361632
Volume17Issue:6Pages:802-810
Abstract

Purpose: Promoters developed for radiogene therapy always show non-negligible transcriptional activities, even when cells are not irradiated. This study developed a tightly radiation-controlled molecular switch based on radiation responsive element (CArG) repeats for in vivo molecular imaging using the Cre/loxP system. Procedures: Different numbers of CArG repeats were cloned as a basal promoter directly, and its pre- and postirradiation transcriptional activities were analyzed by luciferase assay. Nine CArG repeats (E9) were chosen for use as a radiation-controlled molecular switch for the Cre/loxP system, and the feasibility of the switch in vitro and in vivo was demonstrated by luciferase assay and bioluminescence imaging, respectively. Results: The E9 promoter, which exhibits extremely low transcriptional activity, showed a 1.8-fold enhancement after irradiation with a clinical dose of 2 Gy. Both in vitro and in vivo results indicated that E9 is relatively inert but sufficient to trigger the Cre/loxP system. The luciferase activity of stable H1299/pSTOP-FLuc cells transfected with pE9-NLSCre and exposed to 2-Gy radiation can reach 44 % of that of the same cells transfected with pCMV-NLSCre and not subjected to irradiation. By contrast, no appreciable difference was observed in reporter gene expression in both H1299/pSTOPFluc cells and tumors transfected with pE4Pcmv-NLSCre before and after irradiation, because the strong basal transcriptional activity of the CMV promoter, which acts as a copartner of E4, masked the response of E4 to radiation. Conclusions: Our results provide detailed insight into CArG elements as a radiation-controlled molecular switch that can facilitate the development of radiogene therapy.

KeywordCre/loxp System Micropositron Emission Tomography Molecular Switch Radiation-responsive Elements
DOI10.1007/s11307-015-0843-7
URLView the original
Indexed BySCI
Language英语
WOS Research AreaRadiology, Nuclear Medicine & Medical Imaging
WOS SubjectRadiology, Nuclear Medicine & Medical Imaging
WOS IDWOS:000364941700009
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Cited Times [WOS]:1   [WOS Record]     [Related Records in WOS]
Document TypeJournal article
CollectionInstitute of Chinese Medical Sciences
Corresponding AuthorLiu R.-S.
Affiliation1.National Yang-Ming University Taiwan
2.Kaohsiung Medical University
3.Hwa Hsia University of Technology
4.National Comprehensive Mouse Phenotyping and Drug Testing Center
5.National Yang-Ming University, School of Medicine
6.Veterans General Hospital-Taipei
7.Macau University of Science and Technology
Recommended Citation
GB/T 7714
Hsieh Y.-J.,Hwu L.,Ke C.-C.,et al. Demonstration of Tightly Radiation-Controlled Molecular Switch Based on CArG Repeats by In Vivo Molecular Imaging[J]. Molecular Imaging and Biology,2015,17(6):802-810.
APA Hsieh Y.-J..,Hwu L..,Ke C.-C..,Huang A.-L..,Chen F.-D..,...&Liu R.-S..(2015).Demonstration of Tightly Radiation-Controlled Molecular Switch Based on CArG Repeats by In Vivo Molecular Imaging.Molecular Imaging and Biology,17(6),802-810.
MLA Hsieh Y.-J.,et al."Demonstration of Tightly Radiation-Controlled Molecular Switch Based on CArG Repeats by In Vivo Molecular Imaging".Molecular Imaging and Biology 17.6(2015):802-810.
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