Plasmid DNA transfection, revealing tolerant yet separatist mammalian cells
The genome of mammalian cells is challenged by extra-chromosomal DNA introduced by viruses, bacteria, or transfection of plasmid DNA. However, how cells handle these DNA molecules is largely unknown.
We analyzed predominantly Hela cells stably expressing fluorescently tagged LacI protein and transfected with plasmid DNA containing LacO repeats. We found thousands of transfected plasmids in one cytoplasmic cluster being asymmetrically propagated over several divisions. I will present the principles underlying the biased propagation of the plasmid cluster to the cell with the young centrosome and the characteristics of this DNA cluster. Our data suggest that cytoplasmic DNA is maintained in the cytosol of mammalian cells by its enclosing special double membrane that contains LEM-domain proteins and that this cytoplasmic compartment of DNA might act as collector of extra-chromosomal DNA.
ETH Zurich - Institute of Biochemistry, Zurich
Domain 4 - UMR 144 - Subcellular Structure and Cellular Dynamics