Figure 1.

In vivo screening to identify AAV capsids that specifically target the v-SVZ

(A) Schematic illustration of the experimental outline to perform the in vivo screening, including markers used to sort cells of the NSC lineage (see Figure S1 for sorting strategy). IHC of (B) the v-SVZ (scale bar, 50 μm) or (C) the olfactory bulb (OB; scale bars, 200 μm and 30 μm) after injection of library #1 into the lateral ventricle. (D) Mean barcode proportion over all FACS cell types for libraries #1 and #3. Only the 71 capsids shared between the two libraries are shown. (E) Barcode proportion in sample, adjusted for abundance in library (normalized barcode proportion) over all FACS cell types 7 days after library #1 transduction; n = 3 sets per cell type. (F and G) Normalized barcode read count 7 days after library #1 transduction of (F) quiescent NSCs (qNSCs) or of (G) aNSCs; n = 3 sets. (H) Normalized barcode read count over all FACS cell types 7 days after library #3 transduction; n = 2 sets for TAPs and neuroblasts; for all other cell types, n = 3 sets per cell type. (I and J) Normalized barcode read count 7 days after library #3 transduction of (I) qNSCs or of (J) aNSCs; n = 3 sets. (K and L) Normalized barcode read count of AAV2_WT, AAV9_WT, AAV9_A2, and AAV1_P5 after library #1 (K) and #3 (L) transduction of qNSCs, aNSCs, TAPs, neuroblasts, ependymal (Ep) cells, astrocytes, and oligodendrocytes. All mice were 8 weeks old at the time of AAV injection, and all values are given as mean ± SEM. ITR, inverted terminal repeat; BGH, bovine growth hormone poly(A) signal; eYFP, enhanced yellow fluorescent protein; ICV, intracerebroventricular. A set always consists of 6 mice. Three independent experiments were performed resulting in n = 3 sets (3 × 6 mice = 18 mice in total).