Springer Protocols: Abstract: Peptide-Mediated Delivery of Nucleic Acids into Mammalian Cells

Peptide-Mediated Delivery of Nucleic Acids into Mammalian Cells

By: Sébastien Deshayes, Federica Simeoni², May C. Morris², Gilles Divita², Frédéric Heitz²

Abstract

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Control of gene expression using RNA interference (RNAi) technology constitutes a method of choice for investigating gene function in mammalian cells. However, like most oligonucleotide-based strategies, the major limitation of interfering RNA is their poor cellular uptake due to low permeability of the cell membrane to nucleic acids. Several strategies have been developed to improve delivery of oligonucleotides both in cultured cells and in vivo. So far, there is no universal method for their delivery, as they all present several limitations. Peptide-based strategies have been demonstrated to improve the cellular uptake of nucleic acids both in cultured cell and in vivo. This chapter describes a new peptide-based gene delivery system, MPG, which forms stable noncovalent complexes with oligonucleotides and promotes their delivery into a large panel of cell lines without the need for prior chemical covalent coupling. Protocols are described for both adherent and suspension cell lines.

Affiliation(s): (2) Center de Recherches de Biochimie Macromoleculaire, Department of Molecular Biophysics & Thepeutics, Montpellier, France

Book Title: Peptide Characterization and Application Protocols

Series: Methods in Molecular Biology | Volume: 386 | Pub. Date: May-11-2007 | Page Range: 299-308 | DOI: 10.1007/978-1-59745-430-8_11

Subject: Protein Science

Key Words: Peptide carrier - cell-penetrating peptide - cellular uptake - oligonucleotides - siRNA - gene silencing - gene and oligonucleotide delivery - NLS

References

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1.	Luo, D. and Saltzman, M. W. (2000) Synthetic DNA delivery system. Nat. Biotechnol. 18, 33–37.

2.	Niidome, T. and Huang, L. (2002) Gene therapy progress and prospects: non viral vectors. Gene Ther. 10, 991–998.

3.	Rozema, D. B. and Lewis, D. L. (2003) siRNA delivery technologies for mammalian systems. Target 2, 253–260.

4.	Hommel, D. J., Sears, R. M., Georgescu, D., Simmons, D., and Dileone, R. J. (2003) Local gene knockdown in the brain using viral-mediated RNA inteference. Nat. Med. 9, 1539–1543.

5.	Brummelkamp, T. R., Bernards, R., and Agami, R. (2002) Stable suppression of tumorigenicity by virus-mediated RNA interference. Cancer Cell. 2, 243–247.

6.	Xia, H., Mao, Q., Paulson, H. L., and Davidson, B. L. (2002) siRNA-mediated gene silencing in vitro and in vivo. Nat. Biotechnol. 20, 1006–1010.

7.	Simeoni, F., Morris, M. C., Heitz, F., and Divita, G. (2003) Insight into the mechanism of the peptide-based gene delivery system MPG: implication for delivery of siRNA into mammalian cells. Nucleic Acids Res. 31, 2717–2727.

8.	McCaffrey. A. P., Meuse, L., Pham, T. T., Conklin. D. S., Hannon, G. J., and Kay, M. A. (2002) RNA interference in adult mice. Nature 418, 38–39.

9.	Song, E., Lee, S., Wang, J., et al. (2003) RNA interference targeting Fas protects mice from fulminant hepatitis. Nat. Med. 9, 347–351.

10.	Lewis, D. L., Hagstrom, J. E., Loomos, A. G., Wolff, J. A., and Herweijer, H. (2002) Efficient delivery of siRNA for inhibition of gene expression in postnatal mice. Nat. Genet. 32, 107–108.

11.	Gariepy, J. and Kawamura, K. (2000) Vectorial delivery of macromolecules into cells using peptide-based vehicles. Trends Biotechnol. 19, 21–26.

12.	Morris, M. C., Depollier, J., Mery, J., Heitz, F., and Divita, G. (2001) A peptide carrier for the delivery of biologically active proteins into mammalian cells. Nat. Biotechnol. 19, 1173.

13.	Wadia, J. S. and Dowdy, S. F. (2002) Protein transduction technology. Curr. Opin. Biotechnol. 13, 52–56.

14.	Langel, U. ed. (2002) Cell Penetrating Peptides: Processes and Application. CRC, Boca Raton, FL.

15.	Morris, M. C., Chaloin, L., Heitz, F., and Divita, G. (2000) Translocating peptides and proteins and their use for gene delivery. Curr. Opin. Biotechnol. 11, 461–466.

16.	Järver, P. and Langel, U. (2004) The use of cell-penetrating peptides as a tool for gene regulation. Drug Discovery Today 9, 395–402.

17.	Gait, M. J. (2003) Peptide-mediated cellular delivery of antisense oligonucleotides and their analogues. Cell. Mol. Life Sci. 60, 1–10.

18.	Morris, M. C., Vidal, P., Chaloin, L., Heitz, F., and Divita, G. (1997) A new peptide vector for efficient delivery of oligonucleotides into mammalian cells. Nucleic Acids Res. 25, 2730–2736.

19.	Vidal, P., Morris, M. C., Chaloin, L., Heitz, F., and Divita, G. (1997) New strategy for RNA vectorization in mammalian cells. Use of a peptide vector. C R Acad Sci III 320, 279–287.

20.	Morris, M. C., Chaloin, L., Mery, J., Heitz, F., and Divita, G. (1999) A novel potent strategy for gene delivery using a single peptide vector as a carrier. Nucleic Acids Res. 27, 3510–3517.

21.	Morris, M. C., Heitz, F., and Divita G., personal communication.

22.	Marthinet, E., Divita, G., Bernaud, J., Rigal, D., and Baggetto, L. G. (2000) Modulation of the typical multidrug resistance phenotype by targeting the MED-1 region of human MDR1 promoter. Gene Ther. 14, 1224–1233.

23.	Mery, J., Granier, C., Juin, M., and Brugidou J. (1993) Disulfide linkage to polyacrylic resin for automated Fmoc peptide synthesis. Immunochemical applications of peptide resins and mercaptoamide peptides. Int. J. Pept. Protein Res. 42, 44–52.

24.	Vidal, P., Chaloin, L., Mery, J., et al. (1996) Solid-phase synthesis and cellular localization of a C- and/or N-terminal labelled peptide. J. Pept. Sci. 2, 125–133.

25.	Morris, M. C., Chaloin, L., Choob, M., Archdeacon, J., Heitz, F., and Divita, G. (2004) Combination of a new generation of PNAs with a peptide-based carrier enables efficient targeting of cell cycle progression. Gene Ther. 11, 757–764.

26.	Deshayes, S., Plenat, T., Aldrian-Herrada, G., Divita, G., Le Grimellec, C., and Heitz, F. (2004) Primary amphipathic cell-penetrating peptides: structural requirements and interactions with model membranes. Biochemistry 43, 7698–7706.

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