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. 2019 Mar 4;216(3):539–555. doi: 10.1084/jem.20182184

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© 2019 Croft et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms/). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/).

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Figure 1. — Neurons, astrocytes, oligodendrocytes, and microglia in BSCs can be targeted using different rAAV promoter-capsid combinations. (A) Organotypic BSCs were prepared and transduced at 0 DIV with rAAVs (1–2 × 10¹⁰ VGs per well) packaged in capsid serotype 2/1, 2/2, 2/3, 2/4, 2/6, 2/8, and 2/9 to express an EGFP transgene driven by the hCBA promoter. BSCs were maintained in culture until 28 DIV, and then slices were fixed, and fluorescence of EGFP throughout the BSC was imaged. Bar, 100 µm; n = 9 slices from n = 3 wells. (B) BSCs were prepared and transduced at 0 DIV with rAAVs (1–2 × 10¹⁰ VGs per well) with the hCBA promoter to express EGFP packaged in rAAV2/8, a CamKII promoter to express EGFP packaged in rAAV2/8, a CD68 promoter to express EGFP packaged in rAAV2/6 with three mutations—Y731F/Y705F/T492V (TM6)—an MBP promoter to express EGFP packaged in rAAV2/8, and a GFAP promoter to express EBFP packaged in rAAV2/8. BSCs were maintained in culture until 7, 28, and 90 DIV. Slices were then fixed, and fluorescence of EGFP/EBFP of CNS cells at these time points in the BSCs was imaged. Bar, 25 µm.