title: A Novel Ideal Radionuclide Imaging System for Non-invasively Cell Monitoring built on Baculovirus Backbone by Introducing Sleeping Beauty Transposon.
abstract: Sleeping Beauty (SB) transposon is an attractive tool in stable transgene integration both in vitro and in vivo; and we introduced SB transposon into recombinant sodium-iodide symporter baculovirus system (Bac-NIS system) to facilitate long-term expression of recombinant sodium-iodide symporter. In our study, two hybrid baculovirus systems (Bac-eGFP-SB-NeoR and Bac-NIS-SB-NeoR) were successfully constructed and used to infect U87 glioma cells. After G418 selection screening, the Bac-eGFP-SB-NeoR-U87 cells remained eGFP positive, at the 18thand 196thday post transfection (96.03 ± 0.21% and 97.43 ± 0.81%), while eGFP positive population declined significantly at 18 days in cells transfected with unmodified baculovirus construct. NIS gene expression by Bac-NIS-SB-NeoR-U87 cells was also maintained for 28 weeks as determined by radioiodine uptake assay, reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot (WB) assay. When transplanted in mice, Bac-NIS-SB-NeoR-U87 cells also expressed NIS gene stably as monitored by SPECT imaging for 43 days until the tumor-bearing mice were sacrificed. Herein, we showed that incorporation of SB in Bac-NIS system (hybrid Bac-NIS-SB-NeoR) can achieve a long-term transgene expression and can improve radionuclide imaging in cell tracking and monitoring in vivo.
authors.affiliation: Department of Nuclear Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Lv
authors.affiliation: Department of Nuclear Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Pan
authors.affiliation: Department of Nuclear Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
authors.forename: Huijun
authors.initials: H
authors.lastname: Ju
authors.affiliation: Department of Nuclear Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
authors.forename: Jinxin
authors.initials: J
authors.lastname: Zhou
authors.affiliation: Department of Nuclear Medicine, Zhongshan Hospital, Fudan University School of Medicine, Shanghai, China.
authors.forename: Dengfeng
authors.initials: D
authors.lastname: Cheng
authors.affiliation: Department of Nuclear Medicine, Zhongshan Hospital, Fudan University School of Medicine, Shanghai, China.
authors.forename: Hongcheng
authors.initials: H
authors.lastname: Shi
authors.affiliation: Department of Nuclear Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
authors.forename: Yifan
authors.initials: Y
authors.lastname: Zhang
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D045744:Cell Line, Tumor; D058948:Cell Tracking / Q000379:methods*; D004251:DNA Transposable Elements / Q000235:genetics*; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007457:Iodine Radioisotopes / Q000493:pharmacokinetics; D008297:Male; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D011877:Radionuclide Imaging / Q000379:methods*; D015203:Reproducibility of Results; D027981:Symporters / Q000235:genetics* / Q000378:metabolism; D015899:Tomography, Emission-Computed, Single-Photon / Q000379:methods; D019076:Transgenes / Q000235:genetics; D000099209:Sodium-Iodide Symporters
chemical_list: D004251:DNA Transposable Elements; D049452:Green Fluorescent Proteins; D007457:Iodine Radioisotopes; D027981:Symporters; D000099209:Sodium-Iodide Symporters