Retro-Orbital Delivery of AAVs for CNS Wide Astrocyte Targeting

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  • 作者: Domenico Natale, Matthew Holt
  • 杂志: Astrocytes
  • 出版日期: 2025 Mar 21

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摘要

Viral vector-mediated astrocyte targeting in live mice is a popular and valuable method to investigate astrocyte function in the context of intact neural circuits and complex brain physiology. Targeted genetic manipulation and functional investigation of this cell population can be accomplished by utilizing cell type–specific promoters to drive adeno-associated virus (AAV)-mediated transgene expression specifically in astrocytes. Here, we provide a comprehensive protocol for non-invasive retro-orbital (RO) administration of blood–brain barrier (BBB)-crossing AAVs in neonatal and adult mice, such as AAV-PHP.B, AAV-PHP.eB, and AAV.CAP-B22, which results in central nervous system (CNS)-wide transduction. Key procedures outlined include the preparation of AAV solutions for injection, a modified two-handed injection technique for precise and consistent RO injections, and a training strategy to practice mock RO injections using non-toxic dyes. This protocol serves as a valuable resource for researchers interested in exploring the roles of astrocytes in brain functions and neurological disorders.

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作为一家专注于AAV 技术十余年,深耕基因治疗领域的CRO&CDMO,派真生物可提供从载体设计、构建到 AAV、慢病毒和 mRNA 服务的一站式解决方案。凭借深厚的技术实力、卓越的运营管理和高标准的服务交付,我们为全球客户提供一站式CMC解决方案,包括从早期概念验证、成药性评估到IITINDBLA的各个阶段。

 

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