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

• Published in: Neuron (Cambridge, Mass.) Mass.), 2020-08, Vol.107 (4), p.617-630.e6
• Main Authors: 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
• Format: Article
• Language: English
• Subjects: Animals; Biotechnology; Cell division; Cell Lineage; Deoxyribonucleic acid; Design; Development Biology; DNA; DNA vectors; Embryology and Organogenesis; Gene Expression; Gene Transfer Techniques; Genetic engineering; genetic switch; Genetic Vectors; Genomes; genomic integration; Humans; Insertion; Integration; Life Sciences; lineage tracing; Mice; mosaic analysis; Neural Stem Cells; Neurobiology; Neurons and Cognition; Neurosciences; Plasmids; Pluripotency; Regulatory sequences; somatic transgenesis; Stem cell transplantation; Stem cells; Transfection; Transgenes; Transgenic animals; Transposition; transposon systems; Vectors
• ISSN: 0896-6273; 1097-4199
• DOI: 10.1016/j.neuron.2020.05.038

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. [Display omitted] •A gene expression switch powered by genomic integration•Accelerated readout of additive transgenesis with one or multiple vectors•Faithful lineage tracing and mosaic analysis by somatic transfection•Near-universal applicability in cultured cells and animal models Kumamoto et al. introduce iOn, a genetic switch that conditions exogenous transgene expression to integration in the host cell genome by DNA transposition. This system radically simplifies stable transgenesis with one or multiple plasmid vectors, opening new options to genetically manipulate cells in cultured systems and model organisms.