Neural Stem Cell Transduction with AAV

For RNA-seq, NSCs were seeded in 48-well plates (Greiner Bio-One) and incubated overnight. AAV library #1 or library #3 (same libraries as in Weinmann et al.;⁵³ (link) multiplicity of infection [MOI]: 10,000) was added to the media and remained for the duration of 7 days. For IHC, Labtek chambers (Thermo Fisher Scientific) were coated with Poly D-Lysine (PDL; Sigma)/laminin (Sigma), and NSCs were seeded at a density of 2 × 10⁴ cells per square centimeter overnight. AAVs were added (MOI: 10,000) and remained in the media for 1, 3, 5, or 7 days.

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Kremer L.P., Cerrizuela S., Dehler S., Stiehl T., Weinmann J., Abendroth H., Kleber S., Laure A., El Andari J., Anders S., Marciniak-Czochra A., Grimm D, & Martin-Villalba A. (2021). High throughput screening of novel AAV capsids identifies variants for transduction of adult NSCs within the subventricular zone. Molecular Therapy. Methods & Clinical Development, 23, 33-50.

Publication 2021

48-well plates Labtek chambers Poly D-Lysine Sigma)/laminin

Infection Laminin Library Lysine Poly Rna seq

Corresponding Organization : German Cancer Research Center

Other organizations : University Hospital Heidelberg, German Center for Infection Research

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Variable analysis

independent variables

AAV library #1 or library #3
Multiplicity of infection (MOI): 10,000

dependent variables

RNA-seq (gene expression)
Immunohistochemistry (IHC)

control variables

NSCs were seeded in 48-well plates (Greiner Bio-One) and incubated overnight
Labtek chambers (Thermo Fisher Scientific) were coated with Poly D-Lysine (PDL; Sigma)/laminin (Sigma), and NSCs were seeded at a density of 2 × 10^4 cells per square centimeter overnight

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