Direct Readout of Neural Stem Cell Transgenesis with an Integration-Coupled Gene Expression Switch
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.
| Origine | Publication financée par une institution |
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