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- Title
Retrofitting baculoviral vector with <italic>Sleeping Beauty</italic> transposon system: competent for long-term reporter gene imaging in vivo.
- Authors
Pan, Yu; Lv, Jing; Pan, Donghui; Yang, Min; Ju, Huijun; Zhou, Jinxin; Zhu, Liying; Zhang, Yifan
- Abstract
Reporter gene imaging is widely used for non-invasively detecting tumorigenesis, trafficking therapeutic cells, and monitoring treatment effect. Baculoviral vectors (BVs) have been utilized as transgenic vectors in the reporter gene imaging systems in recent years. However, BV-mediated report gene imaging can only provide short-term investigation due to its transient transgene expression, which is incompetent for the long-term applications. In the current study, we reconstructed a series of hybrid BVs with several elements, to investigate the feasibility of this hybrid BV-mediated long-term reporter gene imaging in vivo. We showed that with the indispensable assistance of a positive-selection process, hybrid BV containing <italic>Sleeping Beauty</italic> 100× (SB) transposon system (BV-SB) could significantly prolong the enhanced green fluorescent protein (eGFP) expression for at least 180 days in vitro at nearly 100% eGFP positive percentage and over 1011 arbitrary unit total fluorescence intensity, whereas other hybrid BV-mediated transgene expression gradually faded in only 20 days. Furthermore, BV-SB-mediated eGFP fluorescent reporter gene imaging monitored tumorigenesis in the nude mice for at least 35 days. In addition, we exploited the glucagon-like peptide 1 receptor (<italic>glp-1r</italic>) gene as a radionuclide reporter gene for in vivo micro-PET imaging. At 50th day post-tumor transplantation, the micro-PET imaging showed considerable radiotracer-receptor-binding in vivo, resulted by stable high level of BV-SB-mediated GLP-1R expression in tumor. In summary, we retrofitted BV with the SB transposon system to make it competent for the long-term reporter gene imaging in vivo, which might broaden the application scopes of BV in the long-term molecular imaging and other biomedicine research fields.
- Subjects
BACULOVIRUS genetics; TRANSPOSONS; REPORTER genes; DIAGNOSTIC imaging; GREEN fluorescent protein
- Publication
Applied Microbiology & Biotechnology, 2018, Vol 102, Issue 4, p1933
- ISSN
0175-7598
- Publication type
Article
- DOI
10.1007/s00253-018-8780-4