Retrofitting baculoviral vector with Sleeping Beauty transposon system: competent for long-term reporter gene imaging in vivo
Retrofitting baculoviral vector with Sleeping Beauty transposon system: competent for long-term reporter gene imaging in vivo
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用睡美人转座子系统改造杆状病毒载体：能够进行体内长期报告基因成像
DOI：
10.1007/s00253-018-8780-4
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发表时间：
2018-01
影响因子：
5
通讯作者：
Zhang Yifan
中科院分区：
文献类型：
--
作者：
Pan Yu;Lv Jing;Pan Donghui;Yang Min;Ju Huijun;Zhou Jinxin;Zhu Liying;Zhang Yifan
报告基因成像被广泛应用于无创检测肿瘤发生、转移治疗细胞、监测治疗效果等方面。近年来，杆状病毒载体(BV)被用作报告基因成像系统的转基因载体。然而，BV介导的报告基因成像由于其瞬时转基因表达，只能提供短期的研究，不适合长期应用。在目前的研究中，我们构建了一系列含有多个元件的杂合BV，以探讨这种杂合BV介导的体内长期报告基因成像的可行性。结果表明，在正选择过程的辅助下，含有睡美人100×(SB)转座子系统(BV-SB)的杂交BV在体外可以显著延长增强的绿色荧光蛋白(EGFP)的表达至少180天，几乎100%的EGFP阳性百分率和超过1011个任意单位的总荧光强度，而其他杂交BV介导的转基因表达仅在20天内逐渐消失。此外，BV-SB介导的EGFP荧光报告基因成像至少在35天内监测了裸鼠的肿瘤形成。此外，我们还利用胰升糖素样肽1受体(GLP-1R)基因作为放射性核素报告基因，用于体内Micro-PET成像。在肿瘤移植后第50天，由于BV-SB介导的GLP-1R在肿瘤中的稳定高水平表达，在体内的Micro-PET成像显示了相当大的放射性示踪剂受体结合。综上所述，我们用SB转座子系统对BV进行了改造，使其能够在体内进行长期的报告基因成像，这可能拓宽BV在长期分子成像和其他生物医学研究领域的应用范围。
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 Sleeping Beauty 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 1011arbitrary 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 (glp-1r) 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.
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影响因子：
3.7
作者：
Zhang M;Guo R;Shi S;Miao Y;Zhang Y;Li B
通讯作者：
Li B
影响因子：
3.7
作者：
Pan Y;Liu S;Wu H;Lv J;Xu X;Zhang Y
通讯作者：
Zhang Y
影响因子：
3.8
作者：
Hitchman, Richard B.;Siaterli, Evangelia A.;King, Linda A.
通讯作者：
King, Linda A.
影响因子：
5.1
作者：
Wen-Yi Luo;Yung-Shen Shih;Chie Hung;K. Lo;C. Chiang;Wen-Hsin Lo;S. Huang;S-C Wang;Chenghui Yu;C-H Chien;Y.-C. Hu
通讯作者：
Wen-Yi Luo;Yung-Shen Shih;Chie Hung;K. Lo;C. Chiang;Wen-Hsin Lo;S. Huang;S-C Wang;Chenghui Yu;C-H Chien;Y.-C. Hu
杆状病毒介导的报告基因递送用于有效监测体内胰岛移植的可行性。
影响因子：
3.1
作者：
Lv, Jing;Wu, Haifei;Tian, Jingyan;Zhang, Yifan
通讯作者：
Zhang, Yifan