N-isopropylacrylamide-modified polyethylenimines as effective gene carriers.

25 kDa branched polyethylenimines are modified by N-isopropylacrylamide via Michael addition. An agarose gel retardation assay shows that all derivatives display good binding affinity toward plasmid DNA. The modified PEI-25K shows lower cytotoxicity in MTT assay and better transfection efficiency than unmodified PEI-25K in HeLa cells. The endocytosis efficiency of the optimized complexes is determined to be 99.9% by flow cytometry. More interestingly, although the derivatives are not designed to conjugate with targeting ligands or nuclear localization signals, confocal laser scanning microscopy (CLSM) demonstrates that the optimized derivative results in increased endocytosis and strongly enhanced nuclear uptake compared with PEI-25K.

[1]  Changbai Liu,et al.  Enhancement of TAT cell membrane penetration efficiency by dimethyl sulphoxide. , 2010, Journal of controlled release : official journal of the Controlled Release Society.

[2]  T. W. Johnson,et al.  Preparation and Characterization of Some Derivatives of Poly(ethylenimine) , 1974 .

[3]  T. Reineke,et al.  Hydroxyl stereochemistry and amine number within poly(glycoamidoamine)s affect intracellular DNA delivery. , 2005, Journal of the American Chemical Society.

[4]  Xuesi Chen,et al.  Gene transfection of hyperbranched PEI grafted by hydrophobic amino acid segment PBLG. , 2007, Biomaterials.

[5]  I. Verma,et al.  Gene therapy - promises, problems and prospects , 1997, Nature.

[6]  S. Sahoo,et al.  Intracellular trafficking of nuclear localization signal conjugated nanoparticles for cancer therapy. , 2010, European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences.

[7]  E Marshall,et al.  Gene Therapy Death Prompts Review of Adenovirus Vector , 1999, Science.

[8]  Abdallah,et al.  Gene transfer with lipospermines and polyethylenimines. , 1998, Advanced drug delivery reviews.

[9]  A. Abbott.  Genetic medicine gets real , 2001, Nature.

[10]  F. Wurm,et al.  High-density transfection with HEK-293 cells allows doubling of transient titers and removes need for a priori DNA complex formation with PEI. , 2008, Biotechnology and bioengineering.

[11]  J. Behr,et al.  Gene delivery: a single nuclear localization signal peptide is sufficient to carry DNA to the cell nucleus. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[12]  A. Schätzlein,et al.  PEI-based vesicle-polymer hybrid gene delivery system with improved biocompatibility. , 2004, International journal of pharmaceutics.

[13]  H. Krug,et al.  Peptoidic amino- and guanidinium-carrier systems: targeted drug delivery into the cell cytosol or the nucleus. , 2008, Journal of medicinal chemistry.

[14]  T. Okano,et al.  Temperature-induced intracellular uptake of thermoresponsive polymeric micelles. , 2009, Biomacromolecules.

[15]  Duane D. Miller,et al.  Novel branched poly(ethylenimine)-cholesterol water-soluble lipopolymers for gene delivery. , 2002, Biomacromolecules.

[16]  A. Mesika,et al.  A Regulated, NFκB-Assisted Import of Plasmid DNA into Mammalian Cell Nuclei , 2001 .

[17]  S. W. Kim,et al.  A guanidinylated bioreducible polymer with high nuclear localization ability for gene delivery systems. , 2010, Biomaterials.

[18]  M. Ramezani,et al.  Gene transfer efficiency of high primary amine content, hydrophobic, alkyl-oligoamine derivatives of polyethylenimine. , 2009, Biomaterials.

[19]  W. Tseng,et al.  The role of dextran conjugation in transfection mediated by dextran‐grafted polyethylenimine , 2004, The journal of gene medicine.

[20]  Y. Bae,et al.  Self-assembled polyethylenimine-graft-poly(ε-caprolactone) micelles as potential dual carriers of genes and anticancer drugs , 2007 .

[21]  E. Wagner,et al.  Simple modifications of branched PEI lead to highly efficient siRNA carriers with low toxicity. , 2008, Bioconjugate chemistry.

[22]  R. Mulligan,et al.  The basic science of gene therapy. , 1993, Science.

[23]  A. Klibanov,et al.  Enhancing polyethylenimine's delivery of plasmid DNA into mammalian cells , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[24]  M. D. Brown,et al.  Gene delivery with synthetic (non viral) carriers. , 2001, International journal of pharmaceutics.

[25]  J. Nah,et al.  Degradable polyethylenimine-alt-poly(ethylene glycol) copolymers as novel gene carriers. , 2005, Journal of controlled release : official journal of the Controlled Release Society.

[26]  D. W. Pack,et al.  Partial Acetylation of Polyethylenimine Enhances In Vitro Gene Delivery , 2004, Pharmaceutical Research.

[27]  Lucia Baldi,et al.  A simple high-yielding process for transient gene expression in CHO cells. , 2011, Journal of biotechnology.

[28]  H. Hamm,et al.  Dendritic molecular transporters provide control of delivery to intracellular compartments. , 2007, Bioconjugate chemistry.

[29]  Ye Liu,et al.  Michael addition polymerizations of difunctional amines (AA′) and triacrylamides (B3) , 2006 .

[30]  Y. Bae,et al.  All-trans-retinoic acid (ATRA)-grafted polymeric gene carriers for nuclear translocation and cell growth control. , 2009, Biomaterials.

[31]  F. Wurm,et al.  The Kinetics of Polyethylenimine-Mediated Transfection in Suspension Cultures of Chinese Hamster Ovary Cells , 2008, Molecular biotechnology.

[32]  W. Anderson,et al.  Prospects for in utero human gene therapy. , 1999, Science.

[33]  Peter N. Zawaneh,et al.  Characterizing the structure/function parameter space of hydrocarbon-conjugated branched polyethylenimine for DNA delivery in vitro. , 2006, Journal of controlled release : official journal of the Controlled Release Society.

[34]  Xiabin Jing,et al.  Multi‐armed poly(L‐glutamic acid)‐graft‐oligoethylenimine copolymers as efficient nonviral gene delivery vectors , 2010, The journal of gene medicine.

[35]  Xian‐Zheng Zhang,et al.  A Potential Targeting Gene Vector Based on Biotinylated Polyethyleneimine/Avidin Bioconjugates , 2009, Pharmaceutical Research.

[36]  Y. Durocher,et al.  A simplified polyethylenimine-mediated transfection process for large-scale and high-throughput applications. , 2011, Methods.

[37]  D. Scherman,et al.  A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: polyethylenimine. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[38]  M. Hashida,et al.  In vitro cytotoxicity of macromolecules in different cell culture systems , 1995 .

[39]  S. T. Kim,et al.  Hydrophobic Modification of Polyethyleneimine for Gene Transfectants , 2001 .

[40]  H. Okada,et al.  Enhancement of gene transfection into human dendritic cells using cationic PLGA nanospheres with a synthesized nuclear localization signal. , 2009, International journal of pharmaceutics.