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Journal of Integrative Agriculture  2018, Vol. 17 Issue (08): 1768-1778    DOI: 10.1016/S2095-3119(18)61985-9
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Enhancement of the transfection efficiency of DNA into Crocus sativus L. cells via PEI nanoparticles
Behnam Firoozi, Nasser Zare, Omid Sofalian, Parisa Sheikhzade-Mosadegh
Department of Agronomy and Plant Breeding, Faculty of Agriculture and Natural Resources, University of Mohaghegh Ardabili, Ardabil 56199-11367, Iran
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摘要  Received  22 August, 2017    Accepted  27 January, 2018

Over the past decade, several natural and synthetic cationic polymers have been utilized for gene delivery into cells.  Among them, polyethylenimine (PEI) was used for gene therapy successfully.  The present study investigated the effect of PEI and ultrasound waves on ssDNA delivery into saffron cells.  Gel retardation, dynamic light scattering (DLS) and scanning electron microscopy (SEM) assays were employed to determine the physicochemical properties of PEI/f-DNA  polyplex (complex of PEI and  fluorescently labeled DNA).  Moreover, the cytotoxicity of PEI, PEI/f-DNA polyplex and ultrasound were investigated on saffron cells at different concentrations.  The gel retardation results indicated that the formation and neutralization of the PEI/f-DNA polyplex were completed at N/P=5.  The particle size distribution of the polyplexes was from 50 to 122 nm.  The experimental results revealed that the cytotoxicity of the PEI/f-DNA polyplex was lower than that of PEI alone, hence the cells showed both dose- and exposure duration-dependent responses.  Furthermore, the viability of saffron cells declined extremely after 5 and 10 min sonication but this reduction was not significant at 2 min exposure duration.  The results also indicated that the combined utilization of ultrasound and PEI nanoparticles increased the transfection efficiency of saffron cells up to two times higher than those obtained by PEI or ultrasound separately.
Keywords:  PEI/f-DNA polyplex        plant cell transfection        saffron        sonication  
Received: 22 August 2017   Accepted:
Fund: This study was supported by the University of Mohaghegh Ardabili, India under Grant (51-487).
Corresponding Authors:  Correspondence Nasser Zare, Tel: +98-45-33512081-90, E-mail:    

Cite this article: 

Behnam Firoozi, Nasser Zare, Omid Sofalian, Parisa Sheikhzade-Mosadegh. 2018. Enhancement of the transfection efficiency of DNA into Crocus sativus L. cells via PEI nanoparticles. Journal of Integrative Agriculture, 17(08): 1768-1778.

Ahn H H, Lee M S, Cho M H, Shin Y N, Lee J H, Kim K S, Diamond S L. 2008. DNA/PEI nano-particles for gene delivery of rat bone marrow stem cells. Colloids and Surfaces (A: Physicochemical and Engineering Aspects), 313, 116–120.
Amiji M M (ed). 2004. Polymeric Gene Delivery: Principles and Applications. CRC Press, Florida. USA.
Barampuram S, Zhang Z J. 2011. Recent advances in plant transformation. In: Birchler J, ed. Plant Chromosome Engineering. Methods in Molecular Biology (Methods and Protocols), vol 701. Humana Press, Totowa, New Jersey. USA. pp. 1–35.
Chen Z Y, Qiu R X, Sun X F, Liang K, Zhang J S, Lin Y, Yang F. 2012. The optimization of transfection in MCF-7 cells combining ultrasound irradiation with contrast agent and polyethylenimine. Journal of Cancer Research and Therapeutics, 1, 26.
Cheon S H, Lee K H, Kwon J Y, Choi S H, Song M N, Kim D I. 2009. Enhanced delivery of siRNA complexes by sonoporation in transgenic rice cell suspension cultures. Journal of Microbiology and Biotechnology, 19, 781–786.
Chung Y C, Young T H. 2010. The exhibition of polyethylene imine/DNA coated with oligonucleotides for gene delivery. In: International Conference on Chemistry and Chemical Engineering (ICCCE), IEEE (Institute of Electrical and Electronics Engineers). Kyoto. Japan, pp. 119–123.
Cui J, Cui H, Wang Y, Sun C, Li K, Ren H, Du W. 2012. Application of PEI-modified magnetic nanoparticles as gene transfer vector for the genetic modification of animals. Advances in Materials Science and Engineering, 2012, 1–6.
Deshpande M C, Prausnitz M R. 2007. Synergistic effect of ultrasound and PEI on DNA transfection in vitro. Journal of Controlled Release, 118, 126–135.
Doherty G J, McMahon H T. 2009. Mechanisms of endocytosis. Annual Review of Biochemistry, 78, 857–902.
Fischer R, Stoger E, Schillberg S, Christou P, Twyman R M. 2004. Plant based production of biopharmaceuticals. Current Opinion in Plant Biology, 7, 152–158.
Fu Y Q, Li L H, Wang P W, Qu J, Fu Y P, Wang H, Sun J R. 2012. Delivering DNA into plant cell by gene carriers of ZnS nanoparticles. Chemical Research in Chinese Universities, 28, 672–676.
Godbey W T, Wu K K, Mikos A G. 1999. Size matters: Molecular weight affects the efficiency of poly (ethylenimine) as a gene delivery vehicle. Journal of Biomedical Materials Research, 45, 268–275.
Guzman H R, Nguyen D X, Khan S, Prausnitz M R. 2001. Ultrasound-mediated disruption of cell membranes. II. Heterogeneous effects on cells. The Journal of the Acoustical Society of America, 110, 597–606.
Hamidi M, Azadi A, Rafiei P. 2008. Hydrogel nanoparticles in drug delivery. Advanced Drug Delivery Reviews, 60, 1638–1649.
Karshafian R, Bevan P D, Williams R, Samac S, Burns P N. 2009. Sonoporation by ultrasound-activated microbubble contrast agents: Effect of acoustic exposure parameters on cell membrane permeability and cell viability. Ultrasound in Medicine & Biology, 35, 847–860.
Kim T H, Kim S I, Akaike T, Cho C S. 2005. Synergistic effect of poly (ethylenimine) on the transfection efficiency of galactosylated chitosan/DNA complexes. Journal of Controlled Release, 105, 354–366.
Kim T K, Eberwine J H. 2010. Mammalian cell transfection: The present and the future. Analytical and Bioanalytical Chemistry, 397, 3173–3178.
Liang Y, Liu Z, Shuai X, Wang W, Liu J, Bi W, Wang C, Jing X, Liu Y, Tao E. 2012. Delivery of cationic polymer-siRNA nanoparticles for gene therapies in neural regeneration. Biochemical and Biophysical Research Communications, 421, 690–695.
Louis K S, Siegel A C. 2011. Cell viability analysis using trypan blue: Manual and automated methods. In: Stoddart M, ed., Mammalian Cell Viability. Methods in Molecular Biology (Methods and Protocols), vol 740. Humana Press, USA. pp. 7–12.
McDonald T O, Siccardi M, Moss D, Liptrott N, Giardiello M, Rannard S, Owen A. 2015. The application of nanotechnology to drug delivery in medicine. In: Dolez P, ed., Nano Engineering: Global Approaches to Health and Safety Issues. Elsevier Health Sciences, Toronto, Canada. pp. 173–223.
McKenzie D L, Collard W T, Rice K G. 1999. Comparative gene transfer efficiency of low molecular weight polylysine DNA-condensing peptides. Chemical Biology & Drug Design, 54, 311–318.
Miller D L, Bao S, Morris J E. 1999. Sonoporation of cultured cells in the rotating tube exposure system. Ultrasound in Medicine & Biology, 25, 143–149.
Moghimi S M, Symonds P, Murray J C, Hunter A C, Debska G, Szewczyk A. 2005. A two-stage poly (ethylenimine)-mediated cytotoxicity: Implications for gene transfer/therapy. Molecular Therapy, 11, 990–995.
Mosmann T. 1983. Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. Journal of immunological methods, 65, 55–63.
Murashige T, Skoog F. 1962. A revised medium for rapid growth and bioassays with tobacco cultures. Physiologia Plantarum, 15, 473–497.
Nair R, Varghese S H, Nair B G, Maekawa T, Yoshida Y, Kumar D S. 2010. Nanoparticulate material delivery to plants. Plant Science, 179, 154–163.
Nguyen D N, Green J J, Chan J M, Langer R, Anderson D G. 2009. Polymeric materials for gene delivery and DNA vaccination. Advanced Materials, 21, 847–867.
Ogris M, Steinlein P, Carotta S, Brunner S, Wagner E. 2001. DNA/polyethylenimine transfection particles: Influence of ligands, polymer size, and PEGylation on internalization and gene expression. American Association of Pharmaceutical Scientists, 3, 43–53.
Pasupathy K, Lin S, Hu Q, Luo H, Ke P C. 2008. Direct plant gene delivery with a poly (amidoamine) dendrimer. Biotechnology Journal, 3, 1078–1082.
Phillips L C, Klibanov A L, Wamhoff B R, Hossack J A. 2010. Targeted gene transfection from microbubbles into vascular smooth muscle cells using focused, ultrasound-mediated delivery. Ultrasound in Medicine & Biology, 36, 1470–1480.
Rai M, Bansod S, Bawaskar M, Gade A, dos Santos C A, Seabra A B, Duran N. 2015. Nanoparticles-based delivery systems in plant genetic transformation. In: Nanotechnologies in Food and Agriculture. Springer International Publishing,  Switzerland. pp. 209–239.
Rakoczy-Trojanowska M. 2002. Alternative methods of plant transformation - A short review. Cellular & Molecular Biology Letters, 7, 849–858.
Sanchez-Aguayo I, Rodríguez-Galán J M, García R, Torreblanca J, Pardo J M. 2004. Salt stress enhances xylem development and expression of S-adenosyl-L-methionine synthase in lignifying tissues of tomato plants. Planta, 220, 278–285.
Shih M F, Wu C H, Cherng J Y. 2015. Bioeffects of transient and low-intensity ultrasound on nanoparticles for a safe and efficient DNA delivery. Journal of Nanomedicine & Nanotechnology, 6, 1.
Silva J, Fernandes A R, Baptista P V. 2014. Application of nanotechnology in drug delivery. In: Sezer A D, ed., Application of Nanotechnology in Drug Delivery. InTech, Rijeka, Croatia. pp. 127–154.
Singh S, Singh B K, Yadav S M, Gupta A K. 2015. Applications of nanotechnology in agricultural and their role in disease management. Research Journal of Nanoscience and Nanotechnology, 5, 1–5.
Stumm W, Morgan J J. 1996. Chemical Equilibria and Rates in Natural Waters. Aquatic Chemistry, 1022. John Wiley & Sons, New York.
Sun P, Huang W, Jin M, Wang Q, Fan B, Kang L, Gao Z. 2016. Chitosan-based nanoparticles for survivin targeted siRNA delivery in breast tumor therapy and preventing its metastasis. International Journal of Nanomedicine, 11, 4931–4945.
Sun Y X, Zeng X, Meng Q F, Zhang X Z, Cheng S X, Zhuo R X. 2008. The influence of RGD addition on the gene transfer characteristics of disulfide-containing polyethyleneimine/DNA complexes. Biomaterials, 29, 4356–4365.
Talbot M J, White R G. 2013. Methanol fixation of plant tissue for Scanning Electron Microscopy improves preservation of tissue morphology and dimensions. Plant Methods, 9, 36.
Vain P. 2007. Thirty years of plant transformation technology development. Plant Biotechnology Journal, 5, 221–229.
Vijayanathan V, Thomas T, Thomas T J. 2002. DNA nanoparticles and development of DNA delivery vehicles for gene therapy. Biochemistry, 41, 14085–14094.
Yu K, Zhao J, Yu C, Sun F, Liu Y, Zhang Y, Lee R J, Teng L, Li Y. 2016. Role of four different kinds of polyethylenimines (PEIs) in preparation of polymeric lipid nanoparticles and their anticancer activity study. Journal of Cancer, 7, 872.
Zarnitsyn V G, Prausnitz M R. 2004. Physical parameters influencing optimization of ultrasound-mediated DNA transfection. Ultrasound in Medicine & Biology, 30, 527–538.
Zheng N, Song Z, Liu Y, Yin L, Cheng J. 2017. Gene delivery into isolated Arabidopsis thaliana protoplasts and intact leaves using cationic, α-helical polypeptide. Frontiers of Chemical Science and Engineering, 11, 521–528.
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