title: Charge Type, Charge Spacing, and Hydrophobicity of Arginine-Rich Cell-Penetrating Peptides Dictate Gene Transfection.
abstract: Noncovalent complexation of plasmid DNA (pDNA) with cell-penetrating peptides (CPPs) forms relatively large complexes with poor gene expression. Yet, condensing these CPP-pDNA complexes via addition of calcium chloride produces small and stable nanoparticles with high levels of gene expression. This simple formulation offered high transfection efficiency and negligible cytotoxicity in HEK-293 (a virus-immortalized kidney cell) and A549 (a human lung cancer cell line). Small changes in CPP charge type, charge spacing, and hydrophobicity were studied by using five arginine-rich CPPs: the well-known hydrophilic polyarginine R9 peptide, a hydrophilic RH9 peptide, and three amphiphilic peptides (RA9, RL9, and RW9) with charge distributions that favor membrane penetration. R9 and RW9 nanoparticles were significantly more effective than the other CPPs under most formulation conditions. However, these CPPs exhibit large differences in membrane penetration potential. Maximum transfection resulted from an appropriate balance of complexing with pDNA, releasing DNA, and membrane penetration potential.
authors.affiliation: Department of Pharmaceutical Chemistry, University of Kansas , Lawrence, Kansas 66047, United States.
authors.forename: Nabil A
authors.initials: NA
authors.lastname: Alhakamy
authors.affiliation: Department of Pharmaceutical Chemistry, University of Kansas , Lawrence, Kansas 66047, United States.|Department of Chemical and Petroleum Engineering, University of Kansas , Lawrence, Kansas 66047, United States.
authors.forename: Prajnaparamita
authors.initials: P
authors.lastname: Dhar
authors.affiliation: Department of Pharmaceutical Chemistry, University of Kansas , Lawrence, Kansas 66047, United States.|Department of Chemical and Petroleum Engineering, University of Kansas , Lawrence, Kansas 66047, United States.
authors.forename: Cory J
authors.initials: CJ
authors.lastname: Berkland
mesh_terms: D002118:Calcium / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D057846:Cell-Penetrating Peptides / Q000008:administration & dosage / Q000737:chemistry*; D018014:Gene Transfer Techniques; D057809:HEK293 Cells; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008175:Lung Neoplasms / Q000378:metabolism / Q000473:pathology*; D053758:Nanoparticles / Q000008:administration & dosage / Q000737:chemistry*; D010455:Peptides / Q000008:administration & dosage / Q000737:chemistry*; D011094:Polyethyleneimine / Q000378:metabolism; D014407:Tumor Cells, Cultured
chemical_list: D057846:Cell-Penetrating Peptides; D010455:Peptides; C015462:polyarginine; D011094:Polyethyleneimine; D008156:Luciferases; D002118:Calcium
keywords: amphiphilic; arginine-rich; calcium chloride; cell-penetrating peptides; gene delivery; hydrophilic; nonviral; plasmid DNA; polyarginine; transfection