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

Neuron. 2020 Aug 19;107(4):617-630.e6. doi: 10.1016/j.neuron.2020.05.038. Epub 2020 Jun 18.

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.

Keywords: DNA vectors; genetic engineering; genetic switch; genomic integration; lineage tracing; mosaic analysis; neural stem cells; somatic transgenesis; transposon systems.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Lineage
  • Gene Expression*
  • Gene Transfer Techniques*
  • Genetic Vectors
  • Humans
  • Mice
  • Neural Stem Cells*
  • Transgenes*