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Dual-reporter Imaging and its Potential Application in Tracking Studies

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Abstract

By tracking reporter molecules such as green fluorescent protein and luciferase, researchers can determine physiological status and follow processes both in vitro and in vivo. Here, we describe a dual-reporter imaging method, in which a fusion of eGFP and Luc2 is introduced into hosts using lentiviral particles based on HIV-1. The fusion molecule is both fluorescent and bioluminescent, and is therefore ideal as an optical marker in clinical and research applications. We characterized multiple technical indices of the molecule, including sensibility, biocompatibility, lifetime, and others. Lentiviral particles carrying the reporter were strongly infective in endothelial progenitor (EPC) and GL261 glioma cells, as well as in live mice. By transforming Luc2-eGFP into hosts, morphological and quantitative data can be collected not only from tissue specimens but also from live animal models.

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Acknowledgments

The FUGW vector was kindly provided by Dr. Yanmin Yang at Stanford University. This research was supported by the National Natural Science Foundation of China (81101089 and 81230034), by the Major State Basic Research Development Program of China (2013CB733800), and by Jiangsu Provincial Special Program for Medical Science (BL2013029).

Author Contributions

J. Ding conducted all experiments, integrated data, and edited figures; C. Wang performed the structural analysis and wrote the manuscript; P.C. Li, Z. Zhao, C. Qian, C.X. Wang and Y. Cai provided essential assistance; G.J. Teng directed this study, designed the research and gave key advices.

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Authors and Affiliations

  1. Jiangsu Key Laboratory of Molecular and Functional Imaging, Department of Radiology, Zhongda Hospital, Medical School, Southeast University, #87 Dingjiaqiao Road, Nanjing, 210009, China

    Jie Ding, Zhen Zhao, Cheng Qian, Cong-Xiao Wang, Yu Cai & Gao-Jun Teng

  2. Education Ministry’s Key Laboratory of Developmental Genes and Human Diseases, Institute of Life Sciences, Southeast University, Nanjing, China

    Chao Wang

  3. Department of Interventional Radiology, First Affiliated Hospital of Soochow University, Suzhou, China

    Pei-Cheng Li

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  1. Jie Ding
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Corresponding author

Correspondence to Gao-Jun Teng.

Ethics declarations

This manuscript has not been published or presented elsewhere in part or in entirety, and is not under consideration by another journal. All study participants provided informed consent, and the study design was approved by the appropriate ethics review boards.

Conflict of Interest

All authors discussed the results and approved the manuscript. There are no conflicts of interest to declare.

Electronic supplementary material

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Online Resource 1

Schematic representation of FUGW-Luc2-eGFP. Long terminal repeat, rev response element and woodchuck hepatitis virus posttranscriptional regulatory element were respectively abbreviated as LTR, RRE, and WRE. (GIF 174 kb)

High Resolution (TIFF 107 kb)

Online Resource 2

Structural analysis of eGFP and Luc2. (A) Structure of eGFP in ribbon representation and hydrophobicity surface. (B) Superposition of Luc2 without cofactors (green) and Japanese Genji-botaru (Luciola cruciata) luciferase (magenta) in complex with cofactors AMP and oxyluciferin (yellow). (C) The cofactor-binding pocket in Luciola cruciata luciferase. (GIF 3605 kb)

High Resolution (TIFF 2478 kb)

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Ding, J., Wang, C., Li, PC. et al. Dual-reporter Imaging and its Potential Application in Tracking Studies. J Fluoresc 26, 75–80 (2016). https://doi.org/10.1007/s10895-015-1673-3

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  • DOI: https://doi.org/10.1007/s10895-015-1673-3

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