| Yongwei Shu,Jie Yao,Yang Qu,Jing Zheng,Jing Ding,Lina Zhang,Yefan Wang,Jingyu Zhang,Siqi Tang. Intravenous injection of AAV-PHP.eB across the blood-brain barrier in the adult mouse for central nervous system gene therapy. Oncol Transl Med, 2019, 5: 1-5. |
| 静脉注射AAV-PHP.eB用于中枢神经系统疾病基因治疗的研究 |
| Intravenous injection of AAV-PHP.eB across the blood-brain barrier in the adult mouse for central nervous system gene therapy |
| Received:October 24, 2018 Revised:March 14, 2019 |
| DOI：10.1007/s10330-018-0313-3 |
| 中文关键词: 基因治疗，AAV-PHP.eB，血脑屏障，调节元件，非侵入性病毒注射 |
| 英文关键词: gene therapy; AAV-PHP.eB; blood-brain barrier; regulatory element; noninvasive viral injection |
| 基金项目:哈尔滨市科技人才创新专项资助（2017RAXQJ045）；中央高校基本科研业务费专项资金 |
| Author Name | Affiliation | E-mail | | Yongwei Shu | The Fourth Hospital of Harbin Medical University | Shuyongwei163@163.com | | Jie Yao | East China University of Science and Technology | | | Yang Qu | The Fourth Hospital of Harbin Medical University | | | Jing Zheng | East China University of Science and Technology | | | Jing Ding | The Fourth Hospital of Harbin Medical University | | | Lina Zhang | The Fourth Hospital of Harbin Medical University | | | Yefan Wang | East China University of Science and Technology | | | Jingyu Zhang | The Fourth Hospital of Harbin Medical University | | | Siqi Tang* | East China University of Science and Technology | tangsiqi1@hotmail.com |
|
| Hits: 12149 |
| Download times: 20149 |
| 中文摘要: |
| 摘要：目的 验证成年鼠尾静脉注射AAV-PHP.eB：SYN-GFP后其嗜神经特异性及其跨血脑屏障（blood- brain barrier，BBB）的效率。方法 构建rAAV-SYN-GFP质粒，选择成年C57BL小鼠分别在尾静脉（300nL, 病毒滴度3×109vg） 和脑内前额叶（50L, 病毒滴5×1011vg）注射AAV-PHP.eB：SYN-GFP，在两周时观察脑内GFP信号，四周时观察外周器官中GFP信号。结果 尾静脉注射两周后，在整个脑区显示出GFP表达，特别是在皮质，海马和膝状核中，四周后未在观察外周器官中检测到GFP信号。同时用Western blot进一步确认GFP蛋白仅仅在脑组织中表达；前额叶注射后主要在注射部位观察到GFP信号。
结论AAV-PHP.eB：SYN-GFP可以在成年鼠中有效穿过血脑屏障，同时使用神经元特异性启动子能够让目的基因在神经元中表达。因此AAV-PHP.eB可以作为研究中枢神经疾病的有效载体。 |
| 英文摘要: |
| Objective To verify the neurotypicality of AAV-PHP.eB after tail vein injection in adult mice and its
efficiency in crossing the blood-brain barrier (BBB).
Methods The rAAV-SYN-GFP plasmid was constructed, and adult C57BL mice were injected with AAVPHP.eB: SYN-GFP in the tail vein (300 nL, virus titer 3 × 109 vg) and in the prefrontal lobe (50 L, virus titer
5 × 1011 vg). The green fluorescent protein (GFP) signal in the brain was observed at two weeks, while the
GFP signal in the peripheral organs was observed at four weeks.
Results Two weeks after tail vein injection, GFP expression was observed throughout the brain,
especially in the cortex, hippocampus, and geniculate nucleus. No GFP signal was observed or detected
by western blotting in the peripheral organs after four weeks. GFP signal was observed mainly at the local
site after prefrontal lobe injection.
Conclusion AAV-PHP.eB: SYN-GFP can effectively cross the BBB in adult mice. Using a neuron-specific
promoter allows exogenous gene expression in neurons; therefore, AAV-PHP.eB can be used as an
effective carrier for studying diseases in the central nervous system (CNS). |
|
View Full Text
Download reader HTML全文 |
| Close |