%0 Journal Article %1 schoen_bacterial_2005 %A Schoen, Christoph %A Kolb-Mäurer, Annette %A Geginat, Gernot %A Löffler, Daniela %A Bergmann, Birgit %A Stritzker, Jochen %A Szalay, Aladar A %A Pilgrim, Sabine %A Goebel, Werner %D 2005 %J Cellular Microbiology %K Animals, Cell Cells, Cytosol, Dendritic Epithelial Fluorescent Gene Genetic Green Humans, Line, Listeria Macrophages, Messenger, Mice, Plasmids, Polymerases, Proteins Proteins, Techniques, Transfer Vectors, Viral ag_schoen monocytogenes, {DNA-Directed} {RNA,} {RNA} %N 5 %P 709--724 %R 10.1111/j.1462-5822.2005.00507.x %T Bacterial delivery of functional messenger RNA to mammalian cells %U http://www.ncbi.nlm.nih.gov/pubmed/15839900 %V 7 %X The limited access to the nuclear compartment may constitute one of the major barriers after bacteria-mediated expression plasmid DNA delivery to eukaryotic cells. Alternatively, a self-destructing Listeria monocytogenes strain was used to release translation-competent mRNA directly into the cytosol of epithelial cells, macrophages and human dendritic cells. Enhanced green fluorescent protein (EGFP)-encoding mRNA, adapted for translation in mammalian cells by linking an IRES element to the 5'-end of the egfp coding sequence, was produced by T7 RNA polymerase in the carrier bacteria upon entry into the cytosol where the mRNA is efficiently released from the lysed bacteria and immediately translated in eukaryotic host cells. Besides the much earlier expression of EGFP being detectable already 4 h after infection, the number of EGFP expressing mammalian cells obtained with this novel RNA delivery technique is comparable to or - especially in phagocytic cells - even higher than that obtained with the expression plasmid DNA delivery strategy. Accordingly, bacteria-mediated delivery of ovalbumin-encoding mRNA to macrophages resulted in efficient antigen processing and presentation in vitro indicating that this approach may also be adapted for the in vivo delivery of antigen-encoding mRNA leading to a more efficient immune response when applied to vaccine development.

@article{schoen_bacterial_2005, abstract = {The limited access to the nuclear compartment may constitute one of the major barriers after bacteria-mediated expression plasmid {DNA} delivery to eukaryotic cells. Alternatively, a self-destructing Listeria monocytogenes strain was used to release translation-competent {mRNA} directly into the cytosol of epithelial cells, macrophages and human dendritic cells. Enhanced green fluorescent protein {(EGFP)-encoding} {mRNA,} adapted for translation in mammalian cells by linking an {IRES} element to the 5'-end of the egfp coding sequence, was produced by T7 {RNA} polymerase in the carrier bacteria upon entry into the cytosol where the {mRNA} is efficiently released from the lysed bacteria and immediately translated in eukaryotic host cells. Besides the much earlier expression of {EGFP} being detectable already 4 h after infection, the number of {EGFP} expressing mammalian cells obtained with this novel {RNA} delivery technique is comparable to or - especially in phagocytic cells - even higher than that obtained with the expression plasmid {DNA} delivery strategy. Accordingly, bacteria-mediated delivery of ovalbumin-encoding {mRNA} to macrophages resulted in efficient antigen processing and presentation in vitro indicating that this approach may also be adapted for the in vivo delivery of antigen-encoding {mRNA} leading to a more efficient immune response when applied to vaccine development.}, added-at = {2011-04-07T15:44:20.000+0200}, author = {Schoen, Christoph and {Kolb-Mäurer}, Annette and Geginat, Gernot and Löffler, Daniela and Bergmann, Birgit and Stritzker, Jochen and Szalay, Aladar A and Pilgrim, Sabine and Goebel, Werner}, biburl = {https://www.bibsonomy.org/bibtex/2558f9e7d66706695747303cf0eee53a4/hymi}, doi = {10.1111/j.1462-5822.2005.00507.x}, interhash = {8e3e0d3cbc39e5f343158a510fc3df5b}, intrahash = {558f9e7d66706695747303cf0eee53a4}, issn = {1462-5814}, journal = {Cellular Microbiology}, keywords = {Animals, Cell Cells, Cytosol, Dendritic Epithelial Fluorescent Gene Genetic Green Humans, Line, Listeria Macrophages, Messenger, Mice, Plasmids, Polymerases, Proteins Proteins, Techniques, Transfer Vectors, Viral ag_schoen monocytogenes, {DNA-Directed} {RNA,} {RNA}}, month = may, note = {{PMID:} 15839900}, number = 5, pages = {709--724}, timestamp = {2011-04-07T16:41:09.000+0200}, title = {Bacterial delivery of functional messenger {RNA} to mammalian cells}, url = {http://www.ncbi.nlm.nih.gov/pubmed/15839900}, volume = 7, year = 2005 }