Preparation of Lipid–Peptide–DNA (LPD) Nanoparticles and Their Use for Gene Transfection

  • Fan Zhang
  • Hao-Ying LiEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2118)


Therapeutic gene delivery systems offer the potential for the treatment of a range of inherited and acquired inherited diseases. In contrast with viral gene vectors, the nonviral gene vectors provide a safer alternative and additional advantages such as the improved delivery efficiency, low cost, and often unlimited capacity to package DNA. Here we describe the preparation of a nonviral gene delivery technique based on lipid–peptide–DNA (LPD) complexes. The size of LPD particles is in the nanometer range. The use of these nanoparticulate LPDs results in high efficiency transfections and a high level of gene expression in vitro. LPDs provide a convenient and efficient tool for gene delivery in gene therapy.

Key words

Liposome Nonviral gene vector Gene transfection Flow cytometry 



The manuscript was edited by Mikhail Soloviev.


  1. 1.
    Emery DE (2004) Gene therapy for genetic diseases: on the horizon. Clin Appl Immunol Rev 4:411–422CrossRefGoogle Scholar
  2. 2.
    Hashida M, Nishikawa M, Yamashita F, Takakura Y (2001) Cell-specific delivery of genes with glycosylated carriers. Adv Drug Deliv Rev 52:187–196CrossRefGoogle Scholar
  3. 3.
    Naldini L (2015) Gene therapy returns to Centre stage. Nature 526:351–360CrossRefGoogle Scholar
  4. 4.
    Dunbar CE, High KA, Joung JK et al (2018) Gene therapy comes of age. Science 359:4672CrossRefGoogle Scholar
  5. 5.
    Kay MA, Glorioso JC, Naldini L (2001) Viral vectors for gene therapy: the art of turning infectious agents into vehicles of therapeutics. Nat Med 7:33–40CrossRefGoogle Scholar
  6. 6.
    Russell SJ, Peng K-W, Bell JC (2012) Oncolytic virotherapy. Nat Biotechnol 30:658–670CrossRefGoogle Scholar
  7. 7.
    Huang S, Kamihira M (2013) Development of hybrid viral vectors for gene therapy. Biotechnol Adv 31:208–223CrossRefGoogle Scholar
  8. 8.
    Yin H, Kanasty RL, Eltoukhy AA et al (2014) Non-viral vectors for gene-based therapy. Nat Rev Genet 15:541–555CrossRefGoogle Scholar
  9. 9.
    Wang HY, Jiang YF, Peng HG et al (2015) Recent progress in microRNA delivery for cancer therapy by non-viral synthetic vectors. Adv Drug Deliv Rev 81:142–160CrossRefGoogle Scholar
  10. 10.
    Wang HX, Li M, Lee CM et al (2017) CRISPR/Cas9-based genome editing for disease modeling and therapy: challenges and opportunities for nonviral delivery. Chem Rev 117:9874–9906CrossRefGoogle Scholar
  11. 11.
    Hardee CL, Arevalo-Soliz LM, Hornstein BD et al (2017) Advances in non-viral DNA vectors for gene therapy. Genes (Basel) 8:65CrossRefGoogle Scholar
  12. 12.
    Lin G, Li L, Panwar N et al (2018) Non-viral gene therapy using multifunctional nanoparticles: status, challenges, and opportunities. Coord Chem Rev 374:133–152CrossRefGoogle Scholar
  13. 13.
    Appaiahgari MB, Vrati S (2015) Adenoviruses as gene/vaccine delivery vectors: promises and pitfalls. Expert Opin Biol Ther 15:337–351CrossRefGoogle Scholar
  14. 14.
    Jayant RD, Sosa D, Kaushik A, Atluri V et al (2016) Current status of non-viral gene therapy for CNS disorders. Expert Opin Drug Deliv 13:1433–1445CrossRefGoogle Scholar
  15. 15.
    Zabner J (1997) Cationic lipids used in gene transfer. Adv Drug Deliv Rev 27:17–28CrossRefGoogle Scholar
  16. 16.
    Zhi D, Bai Y, Yang J et al (2018) A review on cationic lipids with different linkers for gene delivery. Adv Colloid Interface Sci 253:114–140CrossRefGoogle Scholar
  17. 17.
    Pedroso de Lima MC, Simões S, Pires P et al (2001) Cationic lipid-DNA complexes in gene delivery: from biophysics to biological applications. Adv Drug Deliv Rev 47:277–294CrossRefGoogle Scholar
  18. 18.
    Morille M, Passirani C, Vonarbourg A et al (2008) Progress in developing cationic vectors for non-viral systemic gene therapy against cancer. Biomaterials 29:3477–3496CrossRefGoogle Scholar
  19. 19.
    Xiang S, Tong H, Shi Q et al (2012) Uptake mechanisms of non-viral gene delivery. J Control Release 158:371–378CrossRefGoogle Scholar
  20. 20.
    Remaut K, Sanders NN, Fayazpour F et al (2006) Influence of plasmid DNA topology on the transfection properties of DOTAP/DOPE lipoplexes. J Control Release 115:335–343CrossRefGoogle Scholar
  21. 21.
    Kim BK, Hwang GB, Seu YB et al (2015) DOTAP/DOPE ratio and cell type determine transfection efficiency with DOTAP-liposomes. BBA-Biomembranes 1848:1996–2001CrossRefGoogle Scholar
  22. 22.
    Zauner W, Farrow NA, Haines AM (2001) In vitro uptake of polystyrene microspheres: effect of particle size, cell line and cell density. J Control Release 71:39–51CrossRefGoogle Scholar
  23. 23.
    Tsai JT, Furstoss KJ, Michnick T et al (2002) Quantitative physical characterization of lipid-polycation-DNA lipoplexes. Biotechnol Appl Biochem 36:13–20CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.Medical Research CouncilLaboratory of Molecular BiologyCambridgeUK
  2. 2.Biomanufacturing Research Centre, School of Mechanical and Electronic EngineeringSoochow UniversitySuzhouChina

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