Research Papers: Imaging

mPlum-IFP 1.4 fluorescent fusion protein may display Förster resonance energy transfer associated properties that can be used for near-infrared based reporter gene imaging

[+] Author Affiliations
Liang-Ting Lin, Bo-Sheng Wang, Jyh-Cheng Chen, Wen-Yi Chang

National Yang-Ming University, Department of Biomedical Imaging and Radiological Sciences, Taipei 112, Taiwan

Chi-Hsien Liu

Chang Gung University, Graduate Institute of Biochemical and Biomedical Engineering, Tao-Yuan 333, Taiwan

Chien Chou

Chang Gung University, Graduate Institute of Electro-Optical Engineering, Taoyuan 33302, Taiwan

Chang Gung University, Health Aging Research Center, Taoyuan 33302, Taiwan

Shu-Jun Chiu

Tzu Chi University, Department of Life Sciences, Hualien 97004, Taiwan

Institute of Radiation Sciences, Tzu Chi Technology College, Hualien 97004, Taiwan

Ren-Shyan Liu

National Yang-Ming University, Department of Biomedical Imaging and Radiological Sciences, Taipei 112, Taiwan

National Yang-Ming University, Biophotonics & Molecular Imaging Research Center, Taipei 112, Taiwan

Taipei Veterans General Hospital, National PET/Cyclotron Center, Department of Nuclear Medicine, Taipei, Taiwan

NRPB/Taiwan Mouse Clinic, Molecular and Genetic Imaging Core, Taipei 112, Taiwan

C. Allen Chang, Yi-Jang Lee

National Yang-Ming University, Department of Biomedical Imaging and Radiological Sciences, Taipei 112, Taiwan

National Yang-Ming University, Biophotonics & Molecular Imaging Research Center, Taipei 112, Taiwan

J. Biomed. Opt. 18(12), 126013 (Dec 16, 2013). doi:10.1117/1.JBO.18.12.126013
History: Received June 4, 2013; Revised October 20, 2013; Accepted November 7, 2013
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Abstract.  Bacteriophytochrome infrared fluorescent protein (IFP) has a long emission wavelength that is appropriate for detecting pathophysiological effects via near-infrared (NIR) based imaging. However, the brightness and photostability of IFP are suboptimal, although an exogenous supply of biliverdin (BV) IXα is able to enhance these properties. In this study, we fused a far red mPlum fluorescent protein to IFP 1.4 via a linker deoxyribonucleic acid (DNA) sequence encoding eight amino acids. The brightness of mPlum-IFP 1.4 fusion protein at the IFP emission channel was comparable to that of native IFP 1.4 protein when fusion protein and IFP 1.4 were excited by 543 and 633 nm using confocal microscopy, respectively. Visualization of IFP 1.4 fluorescence by excitation of mPlum in mPlum-IFP 1.4 fusion protein is likely to be associated with Förster resonance energy transfer (FRET). The FRET phenomenon was also predicted by acceptor photobleaching using confocal microscopy. Furthermore, the expression of mPlum-IFP 1.4 fusion protein could be detected in cell culture and in xenograft tumors in the absence of BV using in vivo imaging system, although the BV was still essential for detecting native IFP 1.4. Therefore, this innovativefluorescent fusion protein would be useful for NIR-based imaging in vitro and in vivo.

Figures in this Article
© 2013 Society of Photo-Optical Instrumentation Engineers

Citation

Liang-Ting Lin ; Bo-Sheng Wang ; Jyh-Cheng Chen ; Chi-Hsien Liu ; Chien Chou, et al.
"mPlum-IFP 1.4 fluorescent fusion protein may display Förster resonance energy transfer associated properties that can be used for near-infrared based reporter gene imaging", J. Biomed. Opt. 18(12), 126013 (Dec 16, 2013). ; http://dx.doi.org/10.1117/1.JBO.18.12.126013


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