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Chemical coupling as a potent strategy for preparation of targeted bacteriophage‐derived gene nanocarriers into eukaryotic cells
Author(s) -
KhalajKondori Mohammad,
Sadeghizadeh Majid,
Behmanesh Mehrdad,
Saggio Isabella,
Monaci Paolo
Publication year - 2011
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.1617
Subject(s) - green fluorescent protein , transfection , microbiology and biotechnology , internalization , nanocarriers , bacteriophage , biology , hek 293 cells , cell culture , chemistry , cell , gene , biochemistry , escherichia coli , genetics , drug , pharmacology
Background The ability to direct efficiently and specifically carriers toward target cells and express the transgene of interest is a critical step in gene therapy trails. The display of targeting molecules on the surface of phage particles might represent a potent solution. In the present study, we evaluated a chemical coupling strategy for displaying human holotransferrin as a targeting molecule on the surface of phage lambda particles for specifically delivering green fluorescent protein (GFP) encoding gene into a human cell line. Methods Human holotransferrin was coupled on the phage lambda particles bearing a GFP‐expression cassette by a chemical coupling strategy to formulate transferrin‐targeted lambda‐GFP (Tf‐targeted‐λ‐GFP) gene nanocarrier. The carrier was then characterized by phage‐enzyme‐linked immunosorbent assay experiments and used for transfection of the human 293T cell line. Particle internalization into the cells was evaluated by immunocytochemical staining and transfection efficacy was studied using fluorescence‐activated cell sorting (FACS) analysis. Results Characterization of the nanocarrier showed a rather high copy number (274 molecules) of transferrin molecules coupled per phage particle. Immunocytochemical staining revealed efficient internalization of the Tf‐targeted‐λ‐GFP compared to wild lambda‐GFP (λ‐GFP) particles. FACS analysis showed 6.72% GFP positive cells for transfections mediated by Tf‐targeted‐λ‐GFP, whereas the value was 0.61% for wild lambda‐GFP particles. Conclusions Our findings highlight chemical coupling as an efficient and straightforward strategy for displaying a targeting molecule at high density on the phage surface, which, in turn, may improve the efficiency of phage‐mediated gene transfer and expression. Copyright © 2011 John Wiley & Sons, Ltd.

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