Specific gene expression in mouse cortical astrocytes is mediated by a 1740bp-GFAP promoter-driven combined adeno-associated virus 2/5/7/8/9

Neuroscience Letters
Xiandong MengWen Jiang

Abstract

We sought to demonstrate the in vivo transduction efficiency and tropism range in astrocytes of a combined-serotype adeno associated virus (AAV2/5/7/8/9). To control expression of enhanced green fluorescent protein (EGFP), a 1740bp glial fibrillary acidic protein (GFAP) promoter was obtained and ligated into vectors of each AAV serotype (2/5/7/8/9). Purified AAVs were then injected into the somatosensory cortex of C57BL/6J mice. Cell-type specific antibodies and subsequent immunofluorescence were used to identify astrocytes (GFAP), neurons (neuronal nuclear antigen, NeuN), microglia (ionized calcium-binding adapter molecule 1, Iba1), and oligodendrocytes (myelin basic protein, MBP), whereby, EGFP expression was measured in each cell type at 1-4 weeks post-injection. Our results indicated that the majority of astrocytes expressed EGFP, while only a small number of neurons expressed EGFP. Both microglia and oligodendrocytes lacked EGFP expression after viral injection. Quantitative analyses revealed that the percentage of EGFP-positive astrocytes was about 98% after viral injection, while the EGFP-positive neuronal percentage was less than 2%. Thus, this study shows that using a combined-serotype AAV carrying a 1740bp GFAP promot...Continue Reading

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Citations

Apr 8, 2016·Journal of Neurophysiology·Yasmine El-ShamaylehGregory D Horwitz
Feb 6, 2017·Journal of Neuroimmune Pharmacology : the Official Journal of the Society on NeuroImmune Pharmacology·Chaitanya R JoshiAnuja Ghorpade
Oct 30, 2020·Frontiers in Immunology·Mathias Linnerbauer, Veit Rothhammer
Jul 3, 2021·Cells·Anastasia A BorodinovaOlga L Vlasova
Aug 13, 2021·Frontiers in Cellular Neuroscience·Aaron J MoulsonPeggy Assinck

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