Engineering nuclear localization signals in modular protein vehicles for gene therapy

Anna Arís, Antonio Villaverde

Research output: Contribution to journalArticleResearchpeer-review

31 Citations (Scopus)

Abstract

Amino acids from 126 to 135 of the SV40 virus T antigen act as efficient nuclear localization signal during infection but also when fused to recombinant proteins. This peptide has been inserted into two alternative acceptor sites of a modified Escherichia coli β-galactosidase which also displays a DNA-binding domain, a cell-binding motif for integrin αvβ3 targeting and cell internalization, and a cryptic nuclear targeting signal naturally present in the bacterial enzyme. In cultured cells, the presence of the SV40 peptide enhances the expression of a delivered DNA up to 30-fold. However, the DNA expression levels are largely depending on the chosen insertion site for the SV40 segment concomitant to the structural impact of peptide accommodation on the protein vehicle. The structural stability of the hybrid protein, apparently critical for efficient gene transfer, is discussed in the context of modular protein engineering to develop non-viral vectors for gene therapy. © 2003 Elsevier Science (USA). All rights reserved.
Original languageEnglish
Pages (from-to)625-631
JournalBiochemical and Biophysical Research Communications
Volume304
Issue number4
DOIs
Publication statusPublished - 16 May 2003

Keywords

  • Gene therapy
  • Molecular interactions
  • Nuclear localization signal
  • SV40 T antigen
  • α β v 3
  • β-Galactosidase

Fingerprint Dive into the research topics of 'Engineering nuclear localization signals in modular protein vehicles for gene therapy'. Together they form a unique fingerprint.

Cite this