Direct Readout of Neural Stem Cell Transgenesis with an Integration-Coupled Gene Expression Switch.
DNA vectors
genetic engineering
genetic switch
genomic integration
lineage tracing
mosaic analysis
neural stem cells
somatic transgenesis
transposon systems
Journal
Neuron
ISSN: 1097-4199
Titre abrégé: Neuron
Pays: United States
ID NLM: 8809320
Informations de publication
Date de publication:
19 08 2020
19 08 2020
Historique:
received:
12
06
2019
revised:
22
04
2020
accepted:
26
05
2020
pubmed:
20
6
2020
medline:
3
11
2020
entrez:
20
6
2020
Statut:
ppublish
Résumé
Stable genomic integration of exogenous transgenes is essential in neurodevelopmental and stem cell studies. Despite tools driving increasingly efficient genomic insertion with DNA vectors, transgenesis remains fundamentally hindered by the impossibility of distinguishing integrated from episomal transgenes. Here, we introduce an integration-coupled On genetic switch, iOn, which triggers gene expression upon incorporation into the host genome through transposition, thus enabling rapid and accurate identification of integration events following transfection with naked plasmids. In vitro, iOn permits rapid drug-free stable transgenesis of mouse and human pluripotent stem cells with multiple vectors. In vivo, we demonstrate faithful cell lineage tracing, assessment of regulatory elements, and mosaic analysis of gene function in somatic transgenesis experiments that reveal neural progenitor potentialities and interaction. These results establish iOn as a universally applicable strategy to accelerate and simplify genetic engineering in cultured systems and model organisms by conditioning transgene activation to genomic integration.
Identifiants
pubmed: 32559415
pii: S0896-6273(20)30407-4
doi: 10.1016/j.neuron.2020.05.038
pmc: PMC7447981
pii:
doi:
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
617-630.e6Informations de copyright
Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of Interests J.L., T.K., R.B.-M., F.M., S.T., K.L., and M. Le have filed a patent application related to the technology described in this article (number EP18305623).
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