Gene transfer into primary human hepatocytes

28 November 2005 - 12:28 Filed in: Publications
hepatology
The article by Boerries Brandenburg, Lars Stockl, Cindy Gutzeit, Martin Roos, Joachim Lupberger, Ruth Schwartlander, Hans Gelderblom, Igor M. Sauer, Peter Hans Hofschneider, Eberhard Hildt in the December issue of HEPATOLOGY reports on a Novel system for efficient gene transfer into primary human hepatocytes via cell permeable HBV virus-like particle. Protein transduction domains (PTD)s have been used to deliver a variety of biologically active cargo across cellular membranes. However the potential of PTDs to mediate transport of nanoparticular structures into the cytoplasm bypassing the endosomal compartment remains unclear. Based on HBV nucleocapsids cell permeable virus like particles (VLP)s harboring a marker gene were established. Cell permeability was achieved by fusion with TLM-PTD. Electron and confocal microscopy revealed that these VLPs translocate as complete particles across the plasma membrane and transverse the cytoplasm towards the nucleus. Inhibition of endocytosis did not affect translocation of these VLPs into the cytoplasm. Based on these particles a gene transfer system was developed. To this end the particles were loaded with DNA encoding SHBs or eGFP that served as marker genes. Using this system for gene transfer in primary human hepatocytes a gene transfer efficiency of ~95% was observed. In conclusion, the TLM-PTD has the potential to mediate efficient transfer of assembled particles and its cargo i.e. nucleic acids into primary human hepatocytes. This provides the basis for development of novel transducible therapeutic or diagnostic particles. Hepatology 2005, 42 (6): 1300-1309