Targeted gene transduction of mammalian cells expressing the HER2/neu receptor by filamentous phage

Lorena Urbanelli, Chiara Ronchini, Laura Fontana, Sylvie Menard, Rosaria Orlandi, Paolo Monaci

Research output: Contribution to journalArticle

41 Citations (Scopus)

Abstract

Screening a random peptide library displayed on phage as fusion to the major capsid protein pVIII identified a ligand binding the human epidermal growth factor receptor 2 (HER2) specifically. By mutating the sequence of this ligand, a "secondary" library was generated, whose panning on HER2-positive cells isolated a phage-borne peptide with increased specific binding to HER2 (phage NL1.1). The same peptide recognised HER2 specifically when expressed as an N-terminal fusion to the minor coat protein pIII. Phage NL1.1 was engineered to include a mammalian expression cassette for a reporter gene within its genome. This modified phage transduced HER2-expressing cells with very high specificity (more than 1000-fold that of parental HER2-negative cells) and with an efficiency comparable to that of chemical transfection protocols. The gene delivery process was remarkably fast, requiring less than 15 minutes incubation of phage with target cells to generate detectable levels of gene expression.

Original languageEnglish
Pages (from-to)965-976
Number of pages12
JournalJournal of Molecular Biology
Volume313
Issue number5
DOIs
Publication statusPublished - Nov 9 2001

Fingerprint

Bacteriophages
Genes
Capsid Proteins
Ligands
Peptide Library
Peptides
Reporter Genes
Libraries
Transfection
human ERBB2 protein
epidermal growth factor receptor-neu receptor
Genome
Gene Expression

Keywords

  • Cancer
  • Gene delivery
  • HER2/neu receptor
  • Phage display
  • Targeting

ASJC Scopus subject areas

  • Virology

Cite this

Targeted gene transduction of mammalian cells expressing the HER2/neu receptor by filamentous phage. / Urbanelli, Lorena; Ronchini, Chiara; Fontana, Laura; Menard, Sylvie; Orlandi, Rosaria; Monaci, Paolo.

In: Journal of Molecular Biology, Vol. 313, No. 5, 09.11.2001, p. 965-976.

Research output: Contribution to journalArticle

Urbanelli, Lorena ; Ronchini, Chiara ; Fontana, Laura ; Menard, Sylvie ; Orlandi, Rosaria ; Monaci, Paolo. / Targeted gene transduction of mammalian cells expressing the HER2/neu receptor by filamentous phage. In: Journal of Molecular Biology. 2001 ; Vol. 313, No. 5. pp. 965-976.
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