Abstract
Methods of noncovalent immobilization of DNA fragments on titanium dioxide nanoparticles (TiO2) were developed to design TiO2∼DNA nanocomposites, which were capable of penetrating through cell membranes. TiO2 nanoparticles of different forms (amorphous, anatase, brookite) with enhanced agglomeration stability were synthesized. The particles were characterized by X-ray diffraction, small-angle X-ray scattering, infrared spectroscopy and atomic force microscopy. Three approaches to the preparation of nanocomposites are described: 1) sorption of polylysine-containing oligonucleotides onto TiO2 nanoparticles, 2) the electrostatic binding of oligonucleotides to TiO2 nanoparticles bearing immobilized polylysine, and 3) sorption of oligonucleotides on TiO2 nanoparticles in the presence of cetyltrimethylammonium bromide (cetavlon). All three methods provide an efficient and stable immobilization of DNA fragments on nanoparticles that leads to nanocomposites with a capacity of up to 40 nmol/mg for an oligonucleotide. DNA fragments in nanocomposites were shown to retain their ability to form complementary complexes. It was demonstrated by confocal laser microscopy that the proposed nanocomposites penetrated into cells without transfection agents and other methods of exposure.
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Abbreviations
- Amf, Anz, and Brt:
-
titanium dioxide nanoparticles in amorphous and crystal (anataze and brookite) forms, respectively
- PA:
-
polyamine
- PL:
-
poly-L-lysine
- PEI:
-
polyethyleneimine
- Sp:
-
spermine
- Sd:
-
spermidine
- XRD:
-
X-ray diffraction
- IR:
-
infrared spectroscopy
- SAXS:
-
small-anlge X-ray scattering
- AFM:
-
atomic-force microscopy
- GE:
-
glycidyl isopropyl ether
- p:
-
phosphate groups of oligonucleotide
- ctab:
-
cetyltrimethylammonium bromide (cetavlon)
- 1× PBS:
-
buffer containing 0.14 M NaCl and 0.01 M K-phosphate
- pH 7.4:
-
Flu, fluorescein residue
- prefix:
-
“d” in designation of oligodeoxynucleotides is omitted
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Original Russian Text © A.S. Levina, Z.R. Ismagilov, M.N. Repkova, N. V. Shikina, S.I. Baiborodin, N.V. Shatskaya, S.N. Zagrebelnyi, V.F. Zarytova, 2013, published in Bioorganicheskaya Khimiya, 2013, Vol. 39, No. 1, pp. 87–98.
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Levina, A.S., Ismagilov, Z.R., Repkova, M.N. et al. Design of TiO2~DNA nanocomposites for penetration into cells. Russ J Bioorg Chem 39, 77–86 (2013). https://doi.org/10.1134/S1068162013010068
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DOI: https://doi.org/10.1134/S1068162013010068