Abstract
The delivery of nucleic acids (NA) like DNA for cell transfection or siRNAs for gene knockdown is of major interest for in vitro studies as well as for applications in vivo. The same is true for other small RNA molecules like miRNAs or miRNA inhibitors (antimiRs). Important nonviral gene delivery vectors include liposomes and cationic polymers. With regard to cationic polymers, polyethylenimines (PEIs) are well established for the delivery of NA, by acting as nanoscale delivery platforms (polyplexes). Their combination with liposomes comprising different phospholipids leads to the formation of lipopolyplexes and can further improve their efficacy and biocompatibility, by combining the favorable properties of lipid systems (high stability, efficient cellular uptake, low cytotoxicity) and PEI (NA condensation, facilitated endosomal release).
In this chapter, optimal lipopolyplex compositions containing different liposomes and certain branched or linear low-molecular weight PEIs are given. This also includes optimal parameters for lipopolyplex generation, based on various PEIs, N/P ratios, lipids, lipid/PEI ratios, and preparation conditions.
Importantly, certain lipopolyplexes retain their biological activity and physicochemical integrity upon prolonged storage at room temperature (RT), in the presence of serum and upon nebulization, thus extending their usefulness toward various applications in vivo.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Piskin E, Dincer S, Turk M (2004) Gene delivery: intelligent but just at the beginning. J Biomater Sci 15(9):1181–1202
Basarkar A, Singh J (2007) Nanoparticulate systems for polynucleotide delivery. Int J Nanomedicine 2(3):353–360
Aigner A (2008) Cellular delivery in vivo of siRNA-based therapeutics. Curr Pharm Des 14(34):3603–3619
Hadinoto K, Sundaresan A, Cheow WS (2013) Lipid-polymer hybrid nanoparticles as a new generation therapeutic delivery platform: a review. Eur J Pharm Biopharm 85 (3 Pt A):427-443.
Torchilin VP (2005) Recent advances with liposomes as pharmaceutical carriers. Nat Rev 4(2):145–160
Maurer N, Fenske DB, Cullis PR (2001) Developments in liposomal drug delivery systems. Expert Opin Biol Ther 1(6):923–947
Landen CN Jr, Chavez-Reyes A, Bucana C, Schmandt R, Deavers MT, Lopez-Berestein G, Sood AK (2005) Therapeutic EphA2 gene targeting in vivo using neutral liposomal small interfering RNA delivery. Cancer Res 65(15):6910–6918
Srinivasan C, Burgess DJ (2009) Optimization and characterization of anionic lipoplexes for gene delivery. J Control Release 136(1):62–70
Chao M, Jiawei X, Zhongxin J, Kuang A (2010) Anionic long-circulating liposomes for delivery of radioiodinated antisense oligonucleotides. Eur J Lipid Sci Technol 112:545–551
Boussif O, Lezoualc’h F, Zanta MA, Mergny MD, Scherman D, Demeneix B, Behr JP (1995) A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: polyethylenimine. Proc Natl Acad Sci U S A 92(16):7297–7301
Neu M, Fischer D, Kissel T (2005) Recent advances in rational gene transfer vector design based on poly(ethylene imine) and its derivatives. J Gene Med 7(8):992–1009
Lai WF (2011) In vivo nucleic acid delivery with PEI and its derivatives: current status and perspectives. Expert Rev Med Devices 8(2):173–185
Hobel S, Aigner A (2013) Polyethylenimines for siRNA and miRNA delivery in vivo. Wiley Interdiscip Rev Nanomed Nanobiotechnol 5(5):484–501
Godbey WT, Wu KK, Mikos AG (1999) Size matters: molecular weight affects the efficiency of poly(ethylenimine) as a gene delivery vehicle. J Biomed Mater Res 45(3):268–275
Tang MX, Szoka FC (1997) The influence of polymer structure on the interactions of cationic polymers with DNA and morphology of the resulting complexes. Gene Ther 4(8):823–832
Lungwitz U, Breunig M, Blunk T, Gopferich A (2005) Polyethylenimine-based non-viral gene delivery systems. Eur J Pharm Biopharm 60(2):247–266
Zuber G, Dauty E, Nothisen M, Belguise P, Behr JP (2001) Towards synthetic viruses. Adv Drug Deliv Rev 52(3):245–253
Behr JP (1997) The proton sponge: a trick to enter cells the viruses did not exploit. Chimia 51:34–36
Godbey WT, Wu KK, Mikos AG (1999) Tracking the intracellular path of poly(ethylenimine)/DNA complexes for gene delivery. Proc Natl Acad Sci U S A 96(9):5177–5181
Pollard H, Remy JS, Loussouarn G, Demolombe S, Behr JP, Escande D (1998) Polyethylenimine but not cationic lipids promotes transgene delivery to the nucleus in mammalian cells. J Biol Chem 273(13):7507–7511
Werth S, Urban-Klein B, Dai L, Hobel S, Grzelinski M, Bakowsky U, Czubayko F, Aigner A (2006) A low molecular weight fraction of polyethylenimine (PEI) displays increased transfection efficiency of DNA and siRNA in fresh or lyophilized complexes. J Control Release 112(2):257–270
Garcia L, Bunuales M, Duzgunes N, Tros de Ilarduya C (2007) Serum-resistant lipopolyplexes for gene delivery to liver tumour cells. Eur J Pharm Biopharm 67(1):58–66
Lee CH, Ni YH, Chen CC, Chou C, Chang FH (2003) Synergistic effect of polyethylenimine and cationic liposomes in nucleic acid delivery to human cancer cells. Biochim Biophys Acta 1611(1-2):55–62
Pelisek J, Gaedtke L, DeRouchey J, Walker GF, Nikol S, Wagner E (2006) Optimized lipopolyplex formulations for gene transfer to human colon carcinoma cells under in vitro conditions. J Gene Med 8(2):186–197
Gaedtke L, Pelisek J, Lipinski KS, Wrighton CJ, Wagner E (2007) Transcriptionally targeted nonviral gene transfer using a beta-catenin/TCF-dependent promoter in a series of different human low passage colon cancer cells. Mol Pharm 4(1):129–139
Hanzlikova M, Soininen P, Lampela P, Mannisto PT, Raasmaja A (2009) The role of PEI structure and size in the PEI/liposome-mediated synergism of gene transfection. Plasmid 61(1):15–21
Schafer J, Hobel S, Bakowsky U, Aigner A (2010) Liposome-polyethylenimine complexes for enhanced DNA and siRNA delivery. Biomaterials 31(26):6892–6900
Ewe A, Schaper A, Barnert S, Schubert R, Temme A, Bakowsky U, Aigner A (2014) Storage stability of optimal liposome-polyethylenimine complexes (lipopolyplexes) for DNA or siRNA delivery. Acta Biomater 10:2663–2673
Ewe A, Aigner A (2014) Nebulization of liposome–polyethylenimine complexes (lipopolyplexes) for DNA or siRNA delivery: Physicochemical properties and biological activity. Eur J Lipid Sci Technol 116(9):1195–1204
Malek A, Czubayko F, Aigner A (2008) PEG grafting of polyethylenimine (PEI) exerts different effects on DNA transfection and siRNA-induced gene targeting efficacy. J Drug Target 16(2):124–139
Urban-Klein B, Werth S, Abuharbeid S, Czubayko F, Aigner A (2005) RNAi-mediated gene-targeting through systemic application of polyethylenimine (PEI)-complexed siRNA in vivo. Gene Ther 12(5):461–466
Chen C, Ridzon DA, Broomer AJ, Zhou Z, Lee DH, Nguyen JT, Barbisin M, Xu NL, Mahuvakar VR, Andersen MR, Lao KQ, Livak KJ, Guegler KJ (2005) Real-time quantification of microRNAs by stem-loop RT-PCR. Nucleic Acids Res 33(20), e179
Acknowledgments
This work was supported by grants from the Saxonian Ministry for Science and Art (Sächsisches Ministerium für Wissenschaft und Kunst, SMWK), the Deutsche Forschungsgemeinschaft (DFG), and the German Cancer Aid (Deutsche Krebshilfe).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2016 Springer Science+Business Media New York
About this protocol
Cite this protocol
Ewe, A., Aigner, A. (2016). Cationic Lipid-Coated Polyplexes (Lipopolyplexes) for DNA and Small RNA Delivery. In: Candiani, G. (eds) Non-Viral Gene Delivery Vectors. Methods in Molecular Biology, vol 1445. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3718-9_12
Download citation
DOI: https://doi.org/10.1007/978-1-4939-3718-9_12
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3716-5
Online ISBN: 978-1-4939-3718-9
eBook Packages: Springer Protocols