Immobilized HIV-1 Tat protein promotes gene transfer via a transactivation-independent mechanism which requires binding of Tat to viral particles

Filomena Nappi, Chiara Chiozzini, Valentina Bordignon, Alessandra Borsetti, Stefania Bellino, Marco Cippitelli, Giovanni Barillari, Antonella Caputo, Mudit Tyagi, Mauro Giacca, Barbara Ensoli

Research output: Contribution to journalArticlepeer-review


Background: Retroviral transduction of cells is improved upon virus adsorption onto immobilized fibronectin (FN) fragments. Because HIV-1 Tat possesses the same functional domains that lead to increased transduction efficiency in FN by colocalization of bound virus and cells, we hypothesized that Tat could enhance gene transfer by a similar mechanism. Methods: Single-cycle replication retro- or lentivirus carrying green fluorescent protein or cloramphenicol acetyltransferase as reporter genes were added to wells coated with Tat or Tat peptides. Wells were extensively washed to remove unbound virus and levels of transduction were detected by measuring reporter gene expression. Virus adsorption to immobilized Tat was measured using a p24 antigen capture assay. Results: Immobilized Tat efficiently binds retro- and lentiviral particles and mediates virus transmission at virus input doses that were otherwise unable to transduce susceptible cells. Virus adsorption to Tat is not mediated by envelope glycoprotein (Env) because immobilized Tat binds and retains vesicular stomatitis virus G (VSV-G) pseudotypes as well as envelope-free particles. HIV-1 Env or VSV-G are required for Tat-assisted transduction, which is abrogated by an antibody blocking the HIV-1 Env-CD4 interaction. Tat-assisted transduction is mediated by the cysteine-rich region of Tat, which is known to be essential for Tat transactivation activity. However, Tat transactivation is not required for Tat-assisted transduction, as indicated by the enhancement of transduction by transactivation-silent Tat mutants. Conclusions: Immobilized Tat promotes virus transduction by a transactivation-independent mechanism, which requires binding of virus to Tat. Recombinant Tat or Tat fragments provide a newmethod to increase efficiency of retro- and lentiviral based gene transfer and gene therapy.

Original languageEnglish
Pages (from-to)955-965
Number of pages11
JournalJournal of Gene Medicine
Issue number11
Publication statusPublished - 2009


  • Gene therapy
  • Immobilized virus
  • Tat
  • Viral transduction

ASJC Scopus subject areas

  • Genetics
  • Molecular Biology
  • Molecular Medicine
  • Genetics(clinical)
  • Drug Discovery


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