Integrin-mediated transfection with peptides containing arginine-glycine-aspartic acid domains

Abstract

Two synthetic peptides comprising an RGD moiety for intecase of the COS-7 and A375M cell lines. With the ECV304 grin binding and a polylysine moiety for DNA binding were cell line, optimum transfection occurred in the absence of tested for transfection efficiency under a variety of different chloroquine. Transfection efficiency of the peptides was conditions. Binding of target cells to the peptide was shown greatest at peptide:DNA ratios of 4:1 (w/w), which were to be strongly dependent on cyclisation of the peptides via calculated to generate complexes containing approxicysteine residues. Low (10 M) concentrations of chloromately 5000 peptide molecules per plasmid. This repquine, added to assist endocytic exit, unexpectedly resented approximately a 6:1 ratio of positive to negative reduced transfection efficiency in two of the cell lines charges. Peptide 5 was shown to have a higher transfectested, COS-7 and ECV304. However, transfection tion efficiency under most conditions, possibly because of efficiency increased at higher chloroquine concentrations more efficient stabilisation of cyclisation by two cysteineand exceeded that in the absence of chloroquine in the cysteine bonds.

Gene transfer with synthetic virus-like particles via the integrin-mediated endocytosis pathway

“…54,55 More closely related work with a vector containing a RGD peptide coupled to an oligolysine showed epithelial cell lines to be transfected; 41 in this system, a minimum number of 5000 ligands per complex, an about 6:1 N/P ratio as well as extra endosomolytic agents such as chloroquine, 42,56 polyethylenimine or inactivated adenovirus 42 were required. In our system, epithelial and fibroblast cells were already efficiently transfected with unmodified PEI-DNA complexes having a strongly positive surface charge via the 'ionic' gene transfer mechanism.…”

The nuclear pore complex is involved in nuclear transfer of plasmid DNA condensed with an oligolysine–RGD peptide containing nuclear localisation properties

“…[32][33][34][35] In addition, we have demonstrated that the presence of a liposome enhances substantially the level of gene expression of this construct. 1,32,36 It was, however, not clear whether this enhancement is entirely due to the targeting function of the oligo-peptide or if it may also be able to confer a nuclear targeting function by virtue of its strong positive charge.…”

Cell-penetrating and cell-targeting peptides in drug delivery

“…Therefore, the RGD peptide was rapidly synthesized in a cyclized form in order to mimic its natural environment within the protein. One of the first "cyclization" was performed by bridging two cysteine side chains placed at the two extremities of the peptide [140,141].…”