A novel method of cell-specific mRNA transfection.
In this study, we developed a cell-specific mRNA transfection system using streptavidin-protein A (ST-PA) fusion protein and monoclonal antibodies (mAbs). We previously reported that ST-PA fusion protein and mAb complexes can transfer certain biotinylated proteins into specific cell types. At this time, we combined an in vitro transcribed biotinylated and self-replicating Sindbis virus genomic RNA with ST-PA fusion protein and mAbs. In the presence of cationic liposomes, to prevent RNA degradation, this complex is able to transfect a reporter gene to specific cancer cells in a mAb does-dependent manner. Even in the absence of cationic liposomes, biotinylated mRNA, ST-PA fusion, and mAb complexes can transfer some types of cancer cell suspension cultures. This cell-specific transfection system is a novel method of introducing various mRNAs into cells that results in high levels of transient protein expression.
Sawai
K
K
Department of Pathology, New York University Medical Center, New York 10016, USA.
Ohno
K
K
Iijima
Y
Y
Levin
B
B
Meruelo
D
D
D000818:Animals; D000911:Antibodies, Monoclonal; D019921:Biotinylation; D015704:CD4 Antigens / Q000276:immunology; D002412:Cations; D002460:Cell Line; D006224:Cricetinae; D018274:Electroporation; D066246:ErbB Receptors / Q000276:immunology; D015316:Genetic Therapy; D006801:Humans; D007074:Immunoglobulin G; D007763:Lac Operon; D008055:Lipids; D008081:Liposomes; D009928:Organ Specificity; D000644:Quaternary Ammonium Compounds; D012333:RNA, Messenger / Q000235:genetics*; D011993:Recombinant Fusion Proteins; D013205:Staphylococcal Protein A; D019809:Streptavidin; D014162:Transfection / Q000379:methods*; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D015704:CD4 Antigens; D002412:Cations; D007074:Immunoglobulin G; D008055:Lipids; D008081:Liposomes; D000644:Quaternary Ammonium Compounds; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D013205:Staphylococcal Protein A; C087276:(3-dimyristyloxypropyl)(dimethyl)(hydroxyethyl)ammonium; D019809:Streptavidin; D066246:ErbB Receptors