Brief intro:
- Author: Domenico Natale, Matthew Holt
- Journal: Astrocytes
- Publication Date: 2025 Mar 21
Abstract
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.
About PackGene
PackGene Biotech is a world-leading CRO and CDMO, excelling in AAV vectors, mRNA, plasmid DNA, and lentiviral vector solutions. Our comprehensive offerings span from vector design and construction to AAV, lentivirus, and mRNA services. With a sharp focus on early-stage drug discovery, preclinical development, and cell and gene therapy trials, we deliver cost-effective, dependable, and scalable production solutions. Leveraging our groundbreaking π-alpha 293 AAV high-yield platform, we amplify AAV production by up to 10-fold, yielding up to 1e+17vg per batch to meet diverse commercial and clinical project needs. Moreover, our tailored mRNA and LNP products and services cater to every stage of drug and vaccine development, from research to GMP production, providing a seamless, end-to-end solution.
