Rational Design of a Triple Reporter Gene for Multimodality Molecular Imaging

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Title:	Rational Design of a Triple Reporter Gene for Multimodality Molecular Imaging
Authors:	Hsieh, Ya-Ju Hwu, Luen Ke, Chien-Chih Yeh, Skye Hsin-Hsien Lin, Chien-Feng Chen, Fu-Du Wang, Hsin-Ell Lin, Kang-Ping Chen, Ran-Chou Liu, Ren-Shyan
Contributors:	教育系
Keywords:	TYPE-1 THYMIDINE KINASE POSITRON-EMISSION-TOMOGRAPHY UBIQUITIN-PROTEASOME SYSTEM FLUORESCENT PROTEINS LIVING SUBJECTS FIREFLY LUCIFERASE EXPRESSION BIOLUMINESCENT NUCLEAR LOCALIZATION
Date:	2014
Issue Date:	2015-01-27 15:45:49 (UTC+8)
Abstract:	Multimodality imaging using noncytotoxic triple fusion (TF) reporter genes is an important application for cell-based tracking, drug screening, and therapy. The firefly luciferase (fl), monomeric red fluorescence protein (mrfp), and truncated herpes simplex virus type 1 thymidine kinase SR39 mutant (ttksr39) were fused together to create TF reporter gene constructs with different order. The enzymatic activities of TF protein in vitro and in vivo were determined by luciferase reporter assay, H-3-FEAU cellular uptake experiment, bioluminescence imaging, and micropositron emission tomography (microPET). The TF construct expressed in H1299 cells possesses luciferase activity and red fluorescence. The tTKSR39 activity is preserved in TF protein and mediates high levels of H-3-FEAU accumulation and significant cell death from ganciclovir (GCV) prodrug activation. In living animals, the luciferase and tTKSR39 activities of TF protein have also been successfully validated by multimodality imaging systems. The red fluorescence signal is relatively weak for in vivo imaging but may expedite FACS-based selection of TF reporter expressing cells. We have developed an optimized triple fusion reporter construct DsRedm-fl-ttksr39 for more effective and sensitive in vivo animal imaging using fluorescence, bioluminescence, and PET imaging modalities, which may facilitate different fields of biomedical research and applications.
Relation:	BIOMED RESEARCH INTERNATIONAL 文獻號碼: 605358
Appears in Collections:	[Department of Education ] journal articles

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