Design and bioinformatics analysis of novel biomimetic peptides as nanocarriers for gene transfer

Document Type : Research Paper

Authors

Department of Nanobiotechnology, Tarbiat Modares University, Tehran, Iran

Abstract

Objective(s):
The introduction of nucleic acids into cells for therapeutic objectives is significantly hindered by the size and charge of these molecules and therefore requires efficient vectors that assist cellular uptake. For several years great efforts have been devoted to the study of development of recombinant vectors based on biological domains with potential applications in gene therapy. Such vectors have been synthesized in genetically engineered approach, resulting in biomacromolecules with new properties that are not present in nature.
Materials and Methods:
In this study, we have designed new peptides using homology modeling with the purpose of overcoming the cell barriers for successful gene delivery through Bioinformatics tools. Three different carriers were designed and one of those with better score through Bioinformatics tools was cloned, expressed and its affinity for pDNA was monitored.
Results:
The resultszz demonstrated that the vector can effectively condense pDNAinto nanoparticles with the average sizes about 100 nm.
Conclusion:
We hope these peptides can overcome the biological barriers associated with gene transfer, and mediate efficient gene delivery.

Keywords


  1. Crystal RG. Transfer of genes to humans: early lessons and obstacles to success. Science. 1995; 270(5235): 404-410.
  2. Pack DW, Hoffman AS, Pun S, Stayton PS. Design and development of polymers for gene delivery. Nat Rev Drug Discov. 2005; 4(7): 581-593.
  3. Somia N, Verma IM. Gene therapy: trials and tribulations. Nat Rev Genet. 2000; 1(2): 91-99.
  4. During MJ. Adeno-associated virus as a gene delivery system. Adv Drug Deliv Rev. 1997; 27(1): 83-94.
  5. Vile RG, Tuszynski A, Castleden S. Retroviral vectors. From laboratory tools to molecular medicine. Mol Biotechnol. 1996; 5(2): 139-158.
  6. Cavazzana-Calvo M, Hacein-Bey S, de Saint Basile G, Gross F, Yvon E, Nusbaum P, et al. Gene therapy of human severe combined immunodeficiency (SCID)-X1 disease. Science. 2000; 288(5466): 669-672.
  7. Sheridan C. Gene therapy finds its niche. Nature biotechnology, 2011. 29(2): 121-128.
  8. Davidson BL, Breakefield XO. Viral vectors for gene delivery to the nervous system. Nat Rev Neurosci. 2003; 4(5): 353-364.
  9. McCarthy HO, Wang Y, Mangipudi SS, Hatefi A. Advances with the use of bio-inspired vectors towards creation of artificial viruses. Expert Opin Drug Deliv. 2010; 7(4): 497-512.
  10. Canine BF, Hatefi A. Development of recombinant cationic polymers for gene therapy research. Adv Drug Deliv Rev. 2010; 62(15): 1524-1529.
  11. Wang Y, Mangipudi SS, Canine BF, Hatefi A. A designer biomimetic vector with a chimeric architecture for targeted gene transfer. J Control Release. 2009; 137(1): 46-53.
  12. Soltani F, Sankian M, Hatefi A, Ramezani M. Development of a novel histone H1-based recombinant fusion peptide for targeted non-viral gene delivery. Int J Pharm. 2013; 441(1-2): 307-315.
  13. Sadeghian F, Hosseinkhani S, Alizadeh A, Hatefi A. Design, engineering and preparation of a multi-domain fusion vector for gene delivery. Int J Pharm. 2012; 427(2): 393-399.
  14. Morris MC, Deshayes S, Heitz F, Divita G. Cell-penetrating peptides: from molecular mechanisms to therapeutics. Biol Cell. 2008; 100(4): 201-217.
  15. Morris MC, Depollier J, Mery J, Heitz F, Divita G. A peptide carrier for the delivery of biologically active proteins into mammalian cells. Nat Biotechnol. 2001; 19(12): 1173-1176.
  16. Zuker M. Mfold web server for nucleic acid folding and hybridization prediction. Nucleic Acids Res. 2003; 31(13): 3406-3415.
  17. Wilkins MR, Gasteiger E, Bairoch A, Sanchez JC, Williams KL, Appel RD, et al. Protein identification and analysis tools in the ExPASy server. Methods Mol Biol. 1999; 112: 531-552.
  18. Zhang Y. I-TASSER server for protein 3D structure prediction. BMC bioinformatics. 2008; 9: 40.
  19. Roy A, Kucukural A, Zhang Y. I-TASSER: a unified platform for automated protein structure and function prediction. Nat Protoc. 2010; 5(4): 725-738.
  20. Morris MC, Vidal P, Chaloin A, Heitz F, Divita G. A new peptide vector for efficient delivery of oligonucleotides into mammalian cells. Nucleic Acids Res. 1997; 25(14): 2730-2736.
  21. Buzon V, Natrajan G, Schibli D, Campelo F, Kozlov MM, Weissenhorn W. Crystal structure of HIV-1 gp41 including both fusion peptide and membrane proximal external regions. PLoS Pathog. 2010; 6(5): e1000880. doi: 10.1371/journal.ppat.1000880.
  22. Veldhoen S, Laufer SD, Trampe A, Restle T. Cellular delivery of small interfering RNA by a non-covalently attached cell-penetrating peptide: quantitative analysis of uptake and biological effect. Nucleic Acids Res. 2006; 34(22): 6561-6573.
  23. Deshayes S, Gerbal-Chaloin S, Morris MC, Aldrian-Herrada G, Charnet P, Divita G, et al. On the mechanism of non-endosomial peptide-mediated cellular delivery of nucleic acids. Biochim Biophys Acta. 2004; 1667(2): 141-147.
  24. Suzuki M, Gerstein M, Johnson T. An NMR study on the DNA-binding SPKK motif and a model for its interaction with DNA. Protein Eng. 1993; 6(6): 565-574.
  25. Khadake JR, Rao MR. Condensation of DNA and chromatin by an SPKK-containing octapeptide repeat motif present in the C-terminus of histone H1. Biochemistry. 1997; 36(5): 1041-1051.