Abstract
In order to study the possibility of using titanium dioxide (TiO2) nanoparticles to deliver peptide nucleic acids (PNA) in eukaryotic cells, a PNA oligomer was synthesized, and a method of PNA immobilization in the form of hybrid DNA/PNA duplexes on the surface of TiO2 nanoparticles covered with polylysine (PL) was developed. The attachment of a DNA/PNA duplex to TiO2 · PL nanoparticles occurs due to electrostatic interactions between the negatively charged DNA chain and the positively charged amino groups of PL. The binding of the PNA to the nanocomposite is achieved through noncovalent Watson-Crick interactions between PNA and complementary DNA. The capacity of the obtained TiO2 · PL · DNA/PNA nano-composites depending on immobilization conditions was 10–30 nmol PNA per 1 mg of TiO2 particles, which corresponds to ∼1–3 PNA molecules per one TiO2 particle with a size of 4–6 nm. It was shown by confocal laser scanning microscopy that fluorescently-labeled PNA molecules in the TiO2 · PL · DNA/FluPNA nano-composites effectively penetrate into HeLa cells without transfection agents, electroporation, or other auxiliary procedures.
Similar content being viewed by others
Abbreviations
- AEEA:
-
aminoethoxyacetate
- Bhoc:
-
benzhydroxycarbonyl
- DIEA:
-
N,N-diisopropyl ethylamine
- FBS:
-
fetal bovine serum
- FITC:
-
fluorescein isothiocyanate
- Flu:
-
fluorescein residue
- Fmoc:
-
9-fluorenylmethoxycarbonyl
- HATU:
-
2-(1-H-7-azabenzotriazole-1-yl)-1, 1,3,3-tetramethyluronium hexafluorophosphate
- IMDM:
-
Iscove’s Modified Dulbecco’s Medium
- NMM:
-
N-methylmorpholine
- PBS:
-
phosphate buffered saline containing 0.01 M KH2PO4, pH 7.5, and 0.14 M NaCl
- TBS:
-
tris buffered saline containing 0.01 M Tris-HCl, pH 7.5, and 0.14 M NaCl
- PS:
-
polystyrene
- PNA:
-
peptide nucleic acids
- FluPNA:
-
fluorescein-labeled PNA
References
Nielsen, P.E., Peptide Nucleic Acids: Protocols and Applications, Wymondam, United Kingdom: Horizon Bioscience, 2004.
Shiraishi, T. and Nielsen, P.E., Methods in Molecular Biology, 2011, vol. 751, pp. 209–221.
Antsypovich, S.I., Usp. Khim., 2002, vol. 71, no. 1, pp. 81–96.
Demidov, V.V., Potaman, V.N., Frank-Kamenetskii, M.D., Egholm, M., Buchard, O., Sönnichsen, S.H., and Nielsen, P.E., Biochem. Pharmacol., 1994, vol. 48, pp. 1310–1313.
Hamilton, S.E., Iyer, M., Norton, J.C., and Corey, D.R., Bioorg. Med. Chem. Lett., 1996, vol. 6, pp. 2897–2900.
Koppelhus, U. and Nielsen, P.E., Adv. Drug Deliv. Rev., 2003, vol. 55, pp. 267–280.
Bogdanenko, E.V., Sviridov, Yu.V., Moskovtsev, A.A., and Zhdanov, R.I., Vopr. Med. Khim., 2000, no. 3, pp. 226–245.
Järver, P. and Langel, U., Biochim. Biophys. Acta, 2006, vol. 1758, pp. 260–263.
Torchilin, V.P., AAPS J., 2007, vol. 9, pp. E128–E147.
Kuznetsova, S.A. and Oretskaya, T.S., Ross. Nanotekhnol., 2010, vol. 5, nos. 9–10, pp. 40–52.
Liua, G., Swierczewskaa, M., Leea, S., and Chena, C., Nano Today, 2010, vol. 5, pp. 524–539.
Torchilin, V.P., Multifunctional Pharmaceutical Nanocarriers, Series: Fundamental Biomedical Technologies, 2008, vol. 4, Part 14.
Cai, R., Hashimoto, K., Itoh, K., Kubota, Y., and Fujishima, A., Bull. Chem. Soc. Jpn., 1991, vol. 64, pp. 1268–1273.
Suzuki, H., Toyooka, T., and Ibuki, Y., Environ. Sci. Technol., 2007, vol. 41, pp. 3018–3024.
Rajh, T., Chen, L.X., Lukas, K., Liu, T., Thurnauer, M.C., and Tiede, D.M., J. Phys. Chem. B, 2002, vol. 106, pp. 10543–10552.
Zhao, D., Chen, C., Wang, Y., Ji, H., Ma, W., Zang, L., and Zhao, J., J. Phys. Chem., 2008, vol. 112, pp. 5993–6001.
Roddick-Lanzilotta, A.D. and McQuillan, A.J., J. Colloid Interface Sci., 1999, vol. 217, pp. 194–202.
Levina, A., Ismagilov, Z., Repkova, M., Shatskaya, N., Shikina, N., Tusikov, F., and Zarytova, V., J. Nanosci. Nanotechnol., 2011, vol. 11, pp. 1–9.
Zarytova, V.F., Zinov’ev, V.V., Ismagilov, Z.R., Levina, A.S., Repkova, M.N., Shikina, N.V., Evdokimov, A.A., Belanov, E.F., Balakhnin, S.M., Serova, O.A., Baiborodin, S.I., Malygin, E.G., and Zagrebel’nyi, S.N., Ross. Nanotekhnol., 2009, vol. 4, pp. 160–163.
Arora, H.C., Jensen, M.P., Yuan, Y., Wu, A., Vogt, S., Paunesku, T., and Woloschak, G.E., Cancer Res., 2012, Jan 18 [Epub ahead of print]. PMID: 22158944
Jeng, H.A. and Swanson, J.J., Environ. Sci. Health A Tox. Hazard Subst. Environ. Eng., 2006, vol. 41, pp. 699–711.
Heinlaan, M., Ivask, A., Blinova, I., Dubourguier, H.C., and Kahru, A., Chemosphere, 2008, vol. 71, pp. 1308–1136.
Liu, H., Ma, L., Zhao, J., Liu, J., Yan, J., Ruan, J., and Hong, F., Biol. Trace Elem. Res., 2009, vol. 129, pp. 170–180.
Brown, E.M., Paunesku, T., Wu, A., Thurn, K.T., Haley, B., Clark, J., Priester, T., and Woloschak, G.E., Anal. Biochem., 2008, vol. 383, pp. 226–235.
Paunesku, T., Rajh, T., Wiederrecht, G., Maser, J., Vogt, S., Stojicevic, N., Protic, M., Lai, B., Oryhon, J., Thurnauer, M., and Woloschak, G., Nat. Mater., 2003, vol. 2, pp. 343–346.
Amirkhanov, N.V., Zhang, K., Aruva, M.R., Thakur, M.L., and Wickstrom, E., Bioconjug. Chem, 2010, vol. 21, pp. 731–740.
Amirkhanov, N., Dimitrov, I., Opitz, A.W., Lai, S., Wagner, N.J., Thakur, M.L., and Wickstrom, E., Biopolymers, 2008, vol. 89, pp. 1061–1076.
Egholm, M., Buchardt, O., Christensen, L., Behrens, C., Freier, S.M., Driver, D.A., Berg, R.H., Kim, S.K., Norden, B., and Nielsen, P.E., Nature, 1993, vol. 365, pp. 556–568.
Giesen, U., Kleider, W., Berding, C., Geiger, A., Orum, H., and Nielsen, P.E., Nucleic Acids Res., 1998, vol. 26, pp. 5004–5006.
Rajh, T., Saponjic, Z., Liu, J., Dimitrijevic, N.M., Scherer, N.F., Vega-Arroyo, M., Zapol, P., Curtiss, L.A., and Thurnauer, M.C., Nano Letters, 2004, vol. 4, pp. 1017–1023.
Kotsokechagia, T., Cellesi, F., Thomas, A., Niederberger, M., and Tirelli, N., Langmuir, 2008, vol. 24, pp. 6988–6997.
Richards, E., Handbook of Biochemistry and Molecular Biology: Nucleic Acids, Fasman, G.D., Ed., Cleavland: CRC Press, 1975, vol. 1, p. 589.
Chan, W.C. and White, P.D., Fmoc Solid Phase Peptide Synthesis. A Practical Approach, New-York: Oxford University Press, 2000.
Perbal, B., A Practical Guide to Molecular Cloning, New York: Academic Press, 1984.
Tadjiki, S., Assemi, S., Deering, C.E., Veranth, J.M., and Miller, J.D., J. Nanopart. Res., 2009, vol. 11, pp. 981–988.
Kaszuba, M., McKnight, D., Connah, M.T., McNeil-Watson, F.K., and Nobbmann, U., J. Nanopart. Res., 2008, vol. 10, pp. 823–829.
Author information
Authors and Affiliations
Corresponding author
Additional information
Original Russian Text © N.V. Amirkhanov, R.N. Amirkhanov, and V.F. Zarytova, 2012, published in Bioorganicheskaya Khimiya, 2012, Vol. 38, No. 6, pp. 692–705.
Rights and permissions
About this article
Cite this article
Amirkhanov, N.V., Amirkhanov, R.N. & Zarytova, V.F. Composites of peptide nucleic acids with ttitanium dioxide nanoparticles. I. Construction of nanocomposites containing DNA/PNA duplexes and their delivery into HeLa cells. Russ J Bioorg Chem 38, 613–624 (2012). https://doi.org/10.1134/S1068162012060027
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1134/S1068162012060027