An approach to increased polyplex gene delivery by peptides selected from a phage display library.
Phage display libraries were screened for peptides to be incorporated in nonviral gene delivery vehicles. Cells in culture were incubated with heptamer random peptide libraries displayed on M13 bacteriophages in three to five copies per phage. Surface-adherent phages were removed or inactivated and the cells were fractionated in a nuclear pellet and supernatant. Bacteriophages from each of the two fractions were amplified and reincubated with the cells. Three successive rounds of selection were performed. Eighteen sequenced clones revealed 14 different sequences. Two sequences were homologous to segments of the HIV gp120 protein. For three sequences, the corresponding synthetic peptides were generated and attached via avidin-biotin to polylysine-condensed plasmid DNA containing a reporter gene. The addition of the peptides led to 8-14 times increase in the expression of the reporter.
Universita' degli Studi di Padova, Dip. Scienze Farmaceutiche, 35131 Padova, Italy.
Margherita
M
Morpurgo
Marc
M
Kirschner
Aurelian
A
Radu
D000595:Amino Acid Sequence; D001360:Avidin / Q000378:metabolism; D017104:Bacteriophage M13 / Q000378:metabolism; D001435:Bacteriophages; D001710:Biotin / Q000378:metabolism; D019921:Biotinylation; D002448:Cell Adhesion; D017931:DNA Primers; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005822:Genetic Vectors; D015699:HIV Envelope Protein gp120 / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D008156:Luciferases / Q000378:metabolism; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry; D011107:Polylysine / Q000737:chemistry / Q000378:metabolism; D011506:Proteins; D014162:Transfection
D017931:DNA Primers; D015699:HIV Envelope Protein gp120; D019151:Peptide Library; D010455:Peptides; D011506:Proteins; D001360:Avidin; D011107:Polylysine; D001710:Biotin; D008156:Luciferases