dc.contributor.author
Zanker, Jeanette
dc.contributor.author
Hüser, Daniela
dc.contributor.author
Savy, Adrien
dc.contributor.author
Lázaro-Petri, Sara
dc.contributor.author
Hammer, Eva-Maria
dc.contributor.author
Schwarzer, Christoph
dc.contributor.author
Heilbronn, Regine
dc.date.accessioned
2024-07-01T12:23:44Z
dc.date.available
2024-07-01T12:23:44Z
dc.identifier.uri
https://refubium.fu-berlin.de/handle/fub188/44037
dc.identifier.uri
http://dx.doi.org/10.17169/refubium-43746
dc.description.abstract
Viral vectors have become important tools for basic research and clinical gene therapy over the past years. However, in vitro testing of vector-derived transgene function can be challenging when specific post-translational modifications are needed for biological activity. Similarly, neuropeptide precursors need to be processed to yield mature neuropeptides. SH-SY5Y is a human neuroblastoma cell line commonly used due to its ability to differentiate into specific neuronal subtypes. In this study, we evaluate the suitability of SH-SY5Y cells in a potency assay for neuropeptide-expressing adeno-associated virus (AAV) vectors. We looked at the impact of neuronal differentiation and compared single-stranded (ss) AAV and self-complementary (sc) AAV transduction at increasing MOIs, RNA transcription kinetics, as well as protein expression and mature neuropeptide production. SH-SY5Y cells proved highly transducible with AAV1 already at low MOIs in the undifferentiated state and even better after neuronal differentiation. Readouts were GFP or neuropeptide mRNA expression. Production of mature neuropeptides was poor in undifferentiated cells. By contrast, differentiated cells produced and sequestered mature neuropeptides into the medium in a MOI-dependent manner.
en
dc.rights.uri
https://creativecommons.org/licenses/by/4.0/
dc.subject
adeno-associated virus
en
dc.subject
gene therapy
en
dc.subject
neuropeptide
en
dc.subject
differentiation
en
dc.subject.ddc
600 Technik, Medizin, angewandte Wissenschaften::610 Medizin und Gesundheit::610 Medizin und Gesundheit
dc.title
Evaluation of the SH-SY5Y cell line as an in vitro model for potency testing of a neuropeptide-expressing AAV vector
dc.type
Wissenschaftlicher Artikel
dcterms.bibliographicCitation.articlenumber
1280556
dcterms.bibliographicCitation.doi
10.3389/fnmol.2023.1280556
dcterms.bibliographicCitation.journaltitle
Frontiers in Molecular Neuroscience
dcterms.bibliographicCitation.originalpublishername
Frontiers Media SA
dcterms.bibliographicCitation.volume
16
refubium.affiliation
Charité - Universitätsmedizin Berlin
refubium.resourceType.isindependentpub
no
dcterms.accessRights.openaire
open access
dcterms.bibliographicCitation.pmid
38098942
dcterms.isPartOf.eissn
1662-5099