Direct Readout of Neural Stem Cell Transgenesis with an Integration-Coupled Gene Expression Switch - Institut Pasteur Access content directly
Journal Articles Neuron Year : 2020

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

Abstract

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.
Fichier principal
Vignette du fichier
Direct Readout of Neural Stem Cell Transgenesis with an Integration.pdf (6 Mo) Télécharger le fichier
Origin : Publication funded by an institution
Loading...

Dates and versions

pasteur-02877084 , version 1 (22-06-2020)

Licence

Attribution - NonCommercial - NoDerivatives

Identifiers

Cite

Takuma Kumamoto, Franck Maurinot, Raphaëlle Barry-Martinet, Célia Vaslin, Sandrine Vandormael-Pournin, et al.. Direct Readout of Neural Stem Cell Transgenesis with an Integration-Coupled Gene Expression Switch. Neuron, 2020, Online ahead of print, pp.S0896-6273(20)30407-4. ⟨10.1016/j.neuron.2020.05.038⟩. ⟨pasteur-02877084⟩
49 View
83 Download

Altmetric

Share

Gmail Facebook X LinkedIn More