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Direct Readout of Neural Stem Cell Transgenesis with an Integration-Coupled Gene Expression Switch

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 tr...

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Autores principales: Kumamoto, Takuma, Maurinot, Franck, Barry-Martinet, Raphaëlle, Vaslin, Célia, Vandormael-Pournin, Sandrine, Le, Mickaël, Lerat, Marion, Niculescu, Dragos, Cohen-Tannoudji, Michel, Rebsam, Alexandra, Loulier, Karine, Nedelec, Stéphane, Tozer, Samuel, Livet, Jean
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cell Press 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7447981/
https://www.ncbi.nlm.nih.gov/pubmed/32559415
http://dx.doi.org/10.1016/j.neuron.2020.05.038
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author Kumamoto, Takuma
Maurinot, Franck
Barry-Martinet, Raphaëlle
Vaslin, Célia
Vandormael-Pournin, Sandrine
Le, Mickaël
Lerat, Marion
Niculescu, Dragos
Cohen-Tannoudji, Michel
Rebsam, Alexandra
Loulier, Karine
Nedelec, Stéphane
Tozer, Samuel
Livet, Jean
author_facet Kumamoto, Takuma
Maurinot, Franck
Barry-Martinet, Raphaëlle
Vaslin, Célia
Vandormael-Pournin, Sandrine
Le, Mickaël
Lerat, Marion
Niculescu, Dragos
Cohen-Tannoudji, Michel
Rebsam, Alexandra
Loulier, Karine
Nedelec, Stéphane
Tozer, Samuel
Livet, Jean
author_sort Kumamoto, Takuma
collection PubMed
description 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.
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spelling pubmed-74479812020-08-31 Direct Readout of Neural Stem Cell Transgenesis with an Integration-Coupled Gene Expression Switch Kumamoto, Takuma Maurinot, Franck Barry-Martinet, Raphaëlle Vaslin, Célia Vandormael-Pournin, Sandrine Le, Mickaël Lerat, Marion Niculescu, Dragos Cohen-Tannoudji, Michel Rebsam, Alexandra Loulier, Karine Nedelec, Stéphane Tozer, Samuel Livet, Jean Neuron Article 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. Cell Press 2020-08-19 /pmc/articles/PMC7447981/ /pubmed/32559415 http://dx.doi.org/10.1016/j.neuron.2020.05.038 Text en © 2020 The Authors http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Article
Kumamoto, Takuma
Maurinot, Franck
Barry-Martinet, Raphaëlle
Vaslin, Célia
Vandormael-Pournin, Sandrine
Le, Mickaël
Lerat, Marion
Niculescu, Dragos
Cohen-Tannoudji, Michel
Rebsam, Alexandra
Loulier, Karine
Nedelec, Stéphane
Tozer, Samuel
Livet, Jean
Direct Readout of Neural Stem Cell Transgenesis with an Integration-Coupled Gene Expression Switch
title Direct Readout of Neural Stem Cell Transgenesis with an Integration-Coupled Gene Expression Switch
title_full Direct Readout of Neural Stem Cell Transgenesis with an Integration-Coupled Gene Expression Switch
title_fullStr Direct Readout of Neural Stem Cell Transgenesis with an Integration-Coupled Gene Expression Switch
title_full_unstemmed Direct Readout of Neural Stem Cell Transgenesis with an Integration-Coupled Gene Expression Switch
title_short Direct Readout of Neural Stem Cell Transgenesis with an Integration-Coupled Gene Expression Switch
title_sort direct readout of neural stem cell transgenesis with an integration-coupled gene expression switch
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7447981/
https://www.ncbi.nlm.nih.gov/pubmed/32559415
http://dx.doi.org/10.1016/j.neuron.2020.05.038
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