title: A versatile targeting system with lentiviral vectors bearing the biotin-adaptor peptide.
abstract: 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.
authors.affiliation: Department of Medicine, David Geffen School of Medicine, University of California, Los Angeles, CA 90095, USA.
authors.forename: Kouki
authors.initials: K
authors.lastname: Morizono
authors.forename: Yiming
authors.initials: Y
authors.lastname: Xie
authors.forename: Gustavo
authors.initials: G
authors.lastname: Helguera
authors.forename: Tracy R
authors.initials: TR
authors.lastname: Daniels
authors.forename: Timothy F
authors.initials: TF
authors.lastname: Lane
authors.forename: Manuel L
authors.initials: ML
authors.lastname: Penichet
authors.forename: Irvin S Y
authors.initials: IS
authors.lastname: Chen
mesh_terms: D001360:Avidin / Q000378:metabolism; D001710:Biotin / Q000378:metabolism*; D019921:Biotinylation; D015153:Blotting, Western; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D008280:Magnetics; D016254:Mutagenesis, Insertional; D010455:Peptides / Q000378:metabolism*; D011990:Receptors, Transferrin / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012845:Sindbis Virus / Q000235:genetics; D014161:Transduction, Genetic; D014759:Viral Envelope Proteins / Q000378:metabolism; D014771:Virion / Q000378:metabolism
chemical_list: D010455:Peptides; D011990:Receptors, Transferrin; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; D001360:Avidin; D001710:Biotin