title: Efficient gene delivery of primary human cells using peptide linked polyethylenimine polymer hybrid.
abstract: Polyethylenimine (PEI) based polymers are efficient agents for cell transfection. However, their use has been hampered due to high cell death associated with transfection thereby resulting in low efficiency of gene delivery within the cells. To circumvent the problem of cellular toxicity, metal binding peptides were linked to PEI. Eight peptide-PEI derivatives were synthesized to improve cell survival and transfection efficiency. TAT linked PEI was used as a control polymer. Peptides linked with PEI amines formed nanogels as shown by electron microscopy and atomic force microscopic measurements. Polymers were characterized by spectroscopic methods and their ability to form complexes with plasmids was tested using electrophoretic studies. These modifications improved polymer biocompatibility as well as cell survival markedly, when compared to PEI alone. A subset of the modified peptide-polymers also showed significantly higher transfection efficiency in primary human cells with respect to the widely used transfection agent, lipofectamine. Study of the underlying mechanism of the observed phenomena revealed lower levels of 'reactive oxygen species' (ROS) in the presence of the peptide-polymers when compared to PEI alone. This was further corroborated with global gene expression analysis which showed upregulation of multiple genes and pathways involved in regulating intracellular oxidative stress.
authors.affiliation: Department of Medicine, Stanford University School of Medicine, Stanford, CA 94305-5111, USA.
authors.forename: Devaveena
authors.initials: D
authors.lastname: Dey
authors.forename: Mohammed
authors.initials: M
authors.lastname: Inayathullah
authors.forename: Andrew S
authors.initials: AS
authors.lastname: Lee
authors.forename: Melburne C
authors.initials: MC
authors.lastname: LeMieux
authors.forename: Xuexiang
authors.initials: X
authors.lastname: Zhang
authors.forename: Yi
authors.initials: Y
authors.lastname: Wu
authors.forename: Divya
authors.initials: D
authors.lastname: Nag
authors.forename: Patricia Eliza
authors.initials: PE
authors.lastname: De Almeida
authors.forename: Leng
authors.initials: L
authors.lastname: Han
authors.forename: Jayakumar
authors.initials: J
authors.lastname: Rajadas
authors.forename: Joseph C
authors.initials: JC
authors.lastname: Wu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D057809:HEK293 Cells; D006801:Humans; D008422:Materials Testing; D008969:Molecular Sequence Data; D015394:Molecular Structure; D018384:Oxidative Stress; D010455:Peptides / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011094:Polyethyleneimine / Q000737:chemistry / Q000378:metabolism; D011108:Polymers / Q000737:chemistry / Q000378:metabolism; D017154:Stromal Cells / Q000166:cytology / Q000502:physiology
chemical_list: D010455:Peptides; D011108:Polymers; D011094:Polyethyleneimine