A universally applicable system for direct readout of eukaryotic cell transgenesis with a novel integration-coupled gene expression switch

Stable integration of exogenous transgenes in the genome of transfected eukaryotic host cells is essential in many biological, biotechnological and biomedical applications.

Naked DNA vectors are increasingly applied for this purpose as an alternative to viral particles due to their low cost, easy production and simplicity of use.

However, transgenesis with these vectors remains fundamentally hindered by the impossibility to distinguish integrated transgenes from those remaining in episomal form, which causes week-long delays to select stable transfected cells with risks of toxicity or epigenetic drift.

We have introduced an efficient strategy to solves these issues, based on a novel genetic switch that conditions DNA transgene expression to incorporation into the host genome.

This integration-coupled On (iOn) switch enables direct and accurate identification of integration events following transfection, opening the way to rapid drug-free stable cell transgenesis.

Here, we propose to develop universally applicable vectors based on this system enabling simplified one-shot stable transfection of eukaryotic cells with one or multiple transgenes, among others.

Our project will set conditions for diffusing a breakthrough technology with strong potential to accelerate genetic engineering in cultured systems and model organisms.