Consistent Delivery of Adeno-Associated Virus via Lateral Tail-Vein Injection in Adult Mice

腺相关病毒 尾静脉 病毒学 病毒 医学 静脉 生物 解剖 内科学 体内 遗传学 重组DNA 基因 载体(分子生物学)
作者
Fady Guirguis,Véronique Bolduc,Matthew J Slarve,Haiyan Zhou,Francesco Muntoni,C. Bönnemann
出处
期刊:Journal of Visualized Experiments [MyJoVE Corporation]
卷期号: (210)
标识
DOI:10.3791/66934
摘要

Many disorders affect multiple organs or involve different regions of the body, so it is critical to deliver therapeutics systemically to target the affected cells located in different sites. Intravenous injection is a widely used systemic delivery route in preclinical studies that assess treatments intended for body-wide administration. In adult mice, it involves the intravenous administration of the therapeutic agent into the mouse's lateral tail veins. When mastered, tail-vein injections are safe and fast, and only require simple and commonly available tools. However, tail-vein injections are technically challenging and require extensive training and continuous practice to ensure the accurate delivery of the intended dose. Here we describe a detailed, optimized, lateral tail-vein injection protocol that we have developed based on our experience and on recommendations that had been previously reported by other groups. Other than the mouse restrainers and insulin syringes, this protocol requires only reagents and equipment that are readily available in most labs. We found that following this protocol results in consistently successful intravenous delivery of adeno-associated virus (AAV) into the tail veins of unsedated 7-9 week-old mice. Additionally, we describe the optimized protocols for the histological detection of fluorescent reporter proteins and vector genome per diploid genome (vg/dg) quantification used to assess AAV transduction and biodistribution. The goal of this protocol is to aid experimenters in easily performing tail-vein injections successfully and consistently, which can reduce the practice time needed to master the technique.

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