Gene transfer via reversible plasmid condensation with cysteine-flanked, internally spaced arginine-rich peptides.
Nonviral gene transfer offers biosafety, stability, and expense advantages over viruses; however, it has suffered from poor efficiency. Because arginine-rich peptides facilitate uptake of macromolecules such as proteins, liposomes, and iron nanoparticles, we explored their potential in enhancing plasmid DNA delivery. In their unmodified form, known protein transduction sequences, including hepta-arginine and Tat(47-57), failed to support effective gene delivery. However, by flanking a core of consecutive arginines with amino- and carboxy-terminal cysteines in vitro gene transfer was observed. Furthermore, interspersing arginines with glycine and histidine residues achieved reversible plasmid condensation and dramatically increased transfection levels in a variety of cell types. Unlike most available cationic homopolymers that function only in vitro, these new peptides also increased gene expression in both murine and human tissue in vivo. Thus, cysteine-flanked, internally spaced arginine-rich (CFIS-R) peptides represent a new approach to efficient nonviral plasmid delivery using rationally designed protein transduction domains.
Siprashvili
Zurab
Z
VA Palo Alto Healthcare System, Palo Alto, CA 94305, USA.
Scholl
Florence A
FA
Oliver
Steven F
SF
Adams
Angie
A
Contag
Christopher H
CH
Wender
Paul A
PA
Khavari
Paul A
PA
D000818:Animals; D001120:Arginine / Q000378:metabolism*; D002522:Chlorocebus aethiops; D003545:Cysteine / Q000235:genetics* / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D005347:Fibroblasts; D015870:Gene Expression; D049452:Green Fluorescent Proteins; D006367:HeLa Cells; D006801:Humans; D008156:Luciferases; D008164:Luminescent Proteins; D051379:Mice; D008856:Microscopy, Fluorescence; D010455:Peptides / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D014162:Transfection / Q000379:methods*
D008164:Luminescent Proteins; D010455:Peptides; D049452:Green Fluorescent Proteins; D004247:DNA; D001120:Arginine; D008156:Luciferases; D003545:Cysteine