Premium
A versatile targeting system with lentiviral vectors bearing the biotin‐adaptor peptide
Author(s) -
Morizono Kouki,
Xie Yiming,
Helguera Gustavo,
Daniels Tracy R.,
Lane Timothy F.,
Penichet Manuel L.,
Chen Irvin S. Y.
Publication year - 2009
Publication title -
the journal of gene medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.689
H-Index - 91
eISSN - 1521-2254
pISSN - 1099-498X
DOI - 10.1002/jgm.1345
Subject(s) - biotinylation , streptavidin , avidin , transduction (biophysics) , biotin , microbiology and biotechnology , fusion protein , chemistry , biology , biochemistry , gene , recombinant dna
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. Copyright © 2009 John Wiley & Sons, Ltd.