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《Oncology and Translational Medicine》 2019年01期
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Intravenous injection of AAV-PHP.eB across the blood-brain barrier in the adult mouse for central nervous system gene therapy

Yongwei Shu  Jie Yao  Yang Qu  Jing Zheng  Jing Ding  Lina Zhang  Yefan Wang  Linlin Zhao  Jingyu Zhang  Siqi Tang  
【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 C57 BL mice were injected with AAV PHP.eB: SYN-GFP in the tail vein(300 nL, virus titer 3 × 10~9 vg) and in the prefrontal lobe(50 L, virus tite5 × 10~(11) 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 loca 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).

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