Background: Targeted gene transduction in vivo is the ultimate preferred method for gene delivery. We previously developed targeting lentiviral vectors that specifically recognize cell surface molecules with conjugated antibodies and mediate targeted gene transduction both in vitro and in vivo. Although effective in some experimental settings, the conjugation of virus with antibodies is mediated by the interaction between protein A and the Fc region of antibodies, which is not as stable as covalent conjugation. We have now developed a more stable conjugation strategy utilizing the interaction between avidin and biotin. Methods: We inserted the biotin-adaptor-peptide, which was biotinylated by secretory biotin ligase at specific sites, into our targeting envelope proteins, enabling conjugation of the pseudotyped virus with avidin, streptavidin or neutravidin. Results: When conjugated with avidin-antibody fusion proteins or the complex of avidin and biotinylated targeting molecules, the vectors could mediate specific transduction to targeted cells recognized by the targeting molecules. When conjugated with streptavidin-coated magnetic beads, transduction by the vectors was targeted to the locations of magnets. Conclusions: This targeting vector system can be used for broad applications of targeted gene transduction using biotinylated targeting molecules or targeting molecules fused with avidin.Fil: Morizono, Kouki. University of California at Los Angeles. School of Medicine; Estados UnidosFil: Xie, Yiming. University of California at Los Angeles. School of Medicine; Estados UnidosFil: Helguera, Gustavo Fernando. University of California at Los Angeles; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Daniels, Tracy R.. University of California at Los Angeles; Estados UnidosFil: Lane T. F.. University of California at Los Angeles. School of Medicine; Estados UnidosFil: Penichet, Manuel L.. University of California at Los Angeles; Estados Unidos. University of California at Los Angeles. School of Medicine; Estados UnidosFil: Chen, Irvin S.Y.. University of California at Los Angeles. School of Medicine; Estados Unido