Stable polyplexes based on arginine-containing oligopeptides for in vivo gene delivery

Abstract

In this study, we investigated to what extent the stability and transduction capacity of polyplexed DNA can be improved by optimizing the condensing peptide sequence. We have synthesized a small library of cationic peptides, at which the lysine/arginine ratio and the cation charge were varied. All peptides were able to compact DNA, at which polyplexes of short lysine-rich sequences were considerably larger than those of elongated or arginine-rich peptides (GM102 and GM202). In addition, the arginine-rich peptides GM102 and GM202 rendered the polyplexes resistant to plasma incubation or DNase I-mediated digestion. While all peptides were found to improve the transfection efficiency in HepG2 cells, only the GM102-and GM202-derived polyplexes could be specifically targeted to HepG2 cells by incorporation of a ligand-derivatized YKAK 8 WK peptide. We propose that GM102 and GM202 combine the advantage of small condensing peptides to give small-sized polyplexes with the superior stability of condensing polymers, which makes GM102 and GM202 excellent candidates for future in vivo gene therapy studies.

Vasopressin V2R-Targeting Peptide Carrier Mediates siRNA Delivery into Collecting Duct Cells

“…Prediction for the structure of the synthetic peptide carrier conjugated with dDAVP showed two separated regions (i.e., ligand domain to V2R and siRNA carrying domain) bisected with a spacer of 4 glycines. The nine-arginine oligopeptide, which is α-helical form, has been known as the arginine-rich cell-penetrating peptide (CPP) for delivering oligonucleotides into the cells [20], [21], [22]. This synthetic peptide carrier made a stable polyplex with siRNA, i.e., siRNA/dDAVP-9r polyplex at the mole ratio over 1∶10 (siRNA:dDAVP-9r).…”

Specific and efficient targeting of adenovirus vectors to macrophages: application of a fusion protein between an adenovirus‐binding fragment and avidin, linked to a biotinylated oligonucleotide

“…The avidin-docking scaffold will allow the use of a whole range of biotinylated ligands including glycosides [49][50][51], peptides [20,[52][53][54][55], entire proteins [29,[56][57][58] or oligonucleotides [44,59,60] to confer affinity for the cell of interest. The affinity of the avidin-biotin dyad, even after attachment of bulky structures, warrants sufficiently tight association between the Ad-binding moiety and the novel homing device and obviates complex coupling reactions and purification steps (for review, see [32]).…”

Bile Acid−Oligopeptide Conjugates Interact with DNA and Facilitate Transfection

“…Twenty-five microliters of a BAC (A6) solution was mixed with 25 μL of pDNA solution containing a variant amount of total pDNA followed by a 30 min incubation. To each 50 μL mixture, 150 μL of serum free (SF) DMEM was added, and 50 μL (triplicate wells) containing ◆ 0.5, □ 1, ▲ 2, and • 3 μg of luciferase plasmid per well was added to subconfluent DAOY cells (1:10 split previous day, ~10 5 cells/well). The cells were exposed to the BAC:pDNA complexes for 3 h, and then the mixture was aspirated, 200 μL of DMEM/10% FBS was added, and cells were outgrown for an additional 48 h. Wells were assayed using the Bright-Glo lysis buffer and Glo-Luciferase assay kit (Promega) as described in the Experimental Section.…”

“…To test the effect of changes in the BAC ratio at different concentrations of plasmid on transfection, 25 μL of BACs was mixed with 25 μL of diluted pDNA followed by a 30 min incubation. To each 50 μL mixture, 150 μL of serum free (SF) DMEM was added, and 50 μL (triplicate wells) containing 0.5, 1, 2, and 3 μg of luciferase plasmid per well was added to DAOY cells, confluent (1:10 split previous day, ~10 5 cells/well). The cells were transfected for 3 h, then the mixture was aspirated and 200 μL of DMEM/10% FBS was added, and the mixture was incubated for 48 h. The cultures were assayed using the Bright-Glo lysis buffer and Glo-Luciferase assay kit (Promega).…”