Isolation and Neuronal Reprogramming of Mouse Embryonic Fibroblasts.
Animals
Cell Culture Techniques
Cell Differentiation
/ genetics
Cell Separation
/ methods
Cellular Reprogramming
/ genetics
Cellular Reprogramming Techniques
Cryopreservation
Fibroblasts
/ cytology
Genetic Vectors
/ biosynthesis
Humans
Mice
Neurogenesis
/ genetics
Neurons
/ cytology
Transcription Factors
/ genetics
Transduction, Genetic
Ascl1
Brn2
Cell fate conversion
Induced neurogenesis
MEF
Myt1l
Neuronal reprogramming
Journal
Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969
Informations de publication
Date de publication:
2021
2021
Historique:
entrez:
29
7
2021
pubmed:
30
7
2021
medline:
1
9
2021
Statut:
ppublish
Résumé
Forced expression of specific neuronal transcription factors in mouse embryonic fibroblasts (MEFs) can lead to their direct conversion into functional neurons. Direct neuronal reprogramming has become a powerful tool to characterize individual factors and molecular mechanisms involved in forced and normal neurogenesis and to generate neuronal cell types for in vitro studies. Here we provide a detailed protocol for the isolation of MEFs devoid of neural tissue and their direct reprogramming into functional neurons by overexpression of neuronal reprogramming factors (Ascl1, Brn2, and Myt1l) using lentiviral vectors. This method enables quick and efficient generation of mouse neurons in vitro for versatile functional and mechanistic characterization.
Identifiants
pubmed: 34324176
doi: 10.1007/978-1-0716-1601-7_1
doi:
Substances chimiques
Transcription Factors
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
1-12Informations de copyright
© 2021. Springer Science+Business Media, LLC, part of Springer Nature.
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