Oncology and Translational Medicine. 2019 Feb
Yongwei Shu, Jie Yao, Yang Qu, Jing Zheng, Jing Ding, Lina Zhang, Yefan Wang, Linlin Zhao, Jingyu Zhang, Siqi Tang
Products used in the paper Details Operation
Plasmid AAV-PHP.eB Plasmid AAV-PHP.eB was purchased from PackGene Biotech, LLC. Request Quote

Research Field: CNS

Keywords: gene therapy; AAV-PHP.eB; blood-brain barrier; regulatory element; noninvasive viral injection

AAV Serotype: AAV-PHP.eB

Dose: virus injection through the tail vein at 5 × 10^11 vg per viral vector. The virus was directly injected into the prefrontal lobe at 3 × 10^9 vg per viral vector.

Routes of Administration: In order to find the vessel, two white mice were injected in the tail vein, while the virus was delivered through the prefrontal lobe in two black mice assisted with a brain stereotaxic apparatus (Olympus, Japan).

Targeted organ: brain

Animal or cell line strain: Virus injections were carried out in 8-week-old male C57BL mice with body weight of 250–300 g.


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).

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