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High throughput screening of novel AAV capsids identifies variants for transduction of adult NSCs within the subventricular zone

The adult mammalian brain entails a reservoir of neural stem cells (NSCs) generating glial cells and neurons. However, NSCs become increasingly quiescent with age, which hampers their regenerative capacity. New means are therefore required to genetically modify adult NSCs for re-enabling endogenous...

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Hauptverfasser: Kremer, Lukas P. M. (VerfasserIn) , Cerrizuela, Santiago (VerfasserIn) , Dehler, Sascha (VerfasserIn) , Stiehl, Thomas (VerfasserIn) , Weinmann, Jonas (VerfasserIn) , Abendroth, Heike (VerfasserIn) , Kleber, Susanne (VerfasserIn) , Laure, Alexander (VerfasserIn) , El Andari, Jihad (VerfasserIn) , Anders, Simon (VerfasserIn) , Marciniak-Czochra, Anna (VerfasserIn) , Grimm, Dirk (VerfasserIn) , Martín-Villalba, Ana (VerfasserIn)
Dokumenttyp: Article (Journal)
Sprache:Englisch
Veröffentlicht: 4 September 2021
In: Molecular therapy. Methods & clinical development
Year: 2021, Jahrgang: 23, Pages: 33-50
ISSN:2329-0501
DOI:10.1016/j.omtm.2021.07.001
Online-Zugang:Verlag, lizenzpflichtig, Volltext: https://doi.org/10.1016/j.omtm.2021.07.001
Verlag, lizenzpflichtig, Volltext: https://www.sciencedirect.com/science/article/pii/S2329050121001182
Volltext
Verfasserangaben:Lukas P.M. Kremer, Santiago Cerrizuela, Sascha Dehler, Thomas Stiehl, Jonas Weinmann, Heike Abendroth, Susanne Kleber, Alexander Laure, Jihad El Andari, Simon Anders, Anna Marciniak-Czochra, Dirk Grimm, and Ana Martin-Villalba
Beschreibung
Zusammenfassung:The adult mammalian brain entails a reservoir of neural stem cells (NSCs) generating glial cells and neurons. However, NSCs become increasingly quiescent with age, which hampers their regenerative capacity. New means are therefore required to genetically modify adult NSCs for re-enabling endogenous brain repair. Recombinant adeno-associated viruses (AAVs) are ideal gene-therapy vectors due to an excellent safety profile and high transduction efficiency. We thus conducted a high-throughput screening of 177 intraventricularly injected barcoded AAV variants profiled by RNA sequencing. Quantification of barcoded AAV mRNAs identified two synthetic capsids, peptide-modified derivative of wild-type AAV9 (AAV9_A2) and peptide-modified derivative of wild-type AAV1 (AAV1_P5), both of which transduce active and quiescent NSCs. Further optimization of AAV1_P5 by judicious selection of the promoter and dose of injected viral genomes enabled labeling of 30%-60% of the NSC compartment, which was validated by fluorescence-activated cell sorting (FACS) analyses and single-cell RNA sequencing. Importantly, transduced NSCs readily produced neurons. The present study identifies AAV variants with a high regional tropism toward the ventricular-subventricular zone (v-SVZ) with high efficiency in targeting adult NSCs, thereby paving the way for preclinical testing of regenerative gene therapy.
Beschreibung:Gesehen am 21.09.2022
Beschreibung:Online Resource
ISSN:2329-0501
DOI:10.1016/j.omtm.2021.07.001

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