Reducible poly(oligo-D-arginine) for enhanced gene expression in mouse lung by intratracheal injection.
Nonarginine (D-R9) has been reported to be one of the most efficacious protein transduction domains (PTDs) for the intracellular cargo delivery such as DNA, RNA, proteins, and particles. Although oligoarginines are capable of forming polyplex with DNA by electrostatic interaction, the length of oligoarginine can affect the toxicity and gene expression. The reducible poly(oligo-D-arginine) (rPOA) composed of the Cys-(D-R9)-Cys repeating unit forming disulfide bonds between terminal cysteinyl-thiol groups of short peptides was hypothesized to show efficient gene transfection without toxicity. The reducible high molecular weight poly(oligo-D-arginine) may fragment into the Cys-(D-R9)-Cys in cellular environments such as cytosol, cell surface, endosomes, and lysosomes, and enhance DNA transfection efficiency. In the present study, in vitro stability, cytotoxicity, and transfection efficiency of DNA/poly(oligo-D-arginine) polyplex were evaluated. In addition, in vivo delivery of DNA into the lung was performed by intratracheal injection of DNA/poly(oligo-D-arginine) polyplex. The in vivo study with rPOA showed higher level of gene expression than PEI, sustaining for 1 week without toxicity. Reducible high molecular weight poly(oligo-D-arginine) based on R9 PTD is a very promising nonviral gene carrier for lung diseases by efficiently condensing, stabilizing, and transfecting DNA.
Won
Young-Wook
YW
Department of Bioengineering, Institute for Bioengineering and Biopharmaceutical Research, Hanyang University, Seoul, Republic of Korea.
Kim
Hyun Ah
HA
Lee
Minhyung
M
Kim
Yong-Hee
YH
D000818:Animals; D002460:Cell Line; D004247:DNA / Q000008:administration & dosage* / Q000009:adverse effects / Q000737:chemistry; D015870:Gene Expression; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D007267:Injections; D008168:Lung / Q000378:metabolism*; D051379:Mice; D009690:Nucleic Acid Conformation; D010455:Peptides / Q000008:administration & dosage* / Q000009:adverse effects / Q000737:chemistry; D051381:Rats; D014161:Transduction, Genetic / Q000379:methods*
D010455:Peptides; C015462:polyarginine; D004247:DNA