DNA passage to nuclei: role of endo-lysosomal circuit in eukaryoticDictyostelium

Abstract

The understanding of DNA passage in eukaryotic cells is still very ambiguous. The route to the nucleus is difficult owing to the barriers, metabolic as well as membranous, posed by the eukaryotic cells. Endocytosis appears to be the most likely process responsible for the transport but is also the major culprit of low transfection efficiencies. Here, we report a study on a eukaryotic amoeba, Dictyostelium discoideum , where by disruption of the endocytic process at the opportune moment, the transformant number increased. We have observed that by disruption of fluid-phase uptake of calcium phosphate DNA nanoparticles, the number of clones increased with the probable increase in number of foreign genes integrating in the host genome. The method described here leads to the possibility of safe and inexpensive methods for transfer of genes required for heterologous recombinant protein production as well as generating therapeutic recombinant cells.