TY - JOUR

T1 - Efficient inhibition of influenza A viral replication in cells by deoxyribozymes delivered by nanocomposites

AU - Repkova, Marina

AU - Levina, Asya

AU - Chelobanov, Boris

AU - Ismagilov, Zinfer

AU - Shatskaya, Natalia

AU - Baiborodin, Sergei

AU - Filippova, Ekaterina

AU - Mazurkova, Natalia

AU - Zarytova, Valentina

N1 - Publisher Copyright: © 2017 Elsevier B.V. and International Society of Chemotherapy

PY - 2017/6/1

Y1 - 2017/6/1

N2 - Nucleic-acid-based drugs are a promising class of novel therapeutics; however, their use in medicine is widely limited because of insufficient delivery into cells. This article proposes a new delivery strategy of nucleic acid fragments into cells as components of TiO2-based nanocomposites. For the first time, unmodified Dz molecules were non-covalently immobilized on TiO2 nanoparticles precovered with polylysine (TiO2•PL) with the formation of (TiO2•PL)•Dz nanocomposites. DNAzymes in the proposed nanocomposites were shown to retain their ability to cleave the RNA target in a cell-free system with the same selectivity as unbound Dz molecules. It was shown by confocal laser microscopy that the fluorescein-labelled (TiO2•PL)•DzFlu nanocomposites penetrate into eukaryotic cells, where DzFlu is internalized in the cytoplasm and predominantly in nuclei. Delivery of deoxyribozymes into cells in the proposed nanocomposites permits very efficient interactions with RNA targets inside cells. This was demonstrated by an example of inhibition of H5N1 influenza A virus replication (inhibition by a factor of ca. 3000). This effect was one order of magnitude higher than with using lipofectamine as the transfection agent. The proposed (TiO2•PL)•Dz nanocomposites demonstrated high antiviral activity and are thus potent as nucleic-acid-based drugs.

AB - Nucleic-acid-based drugs are a promising class of novel therapeutics; however, their use in medicine is widely limited because of insufficient delivery into cells. This article proposes a new delivery strategy of nucleic acid fragments into cells as components of TiO2-based nanocomposites. For the first time, unmodified Dz molecules were non-covalently immobilized on TiO2 nanoparticles precovered with polylysine (TiO2•PL) with the formation of (TiO2•PL)•Dz nanocomposites. DNAzymes in the proposed nanocomposites were shown to retain their ability to cleave the RNA target in a cell-free system with the same selectivity as unbound Dz molecules. It was shown by confocal laser microscopy that the fluorescein-labelled (TiO2•PL)•DzFlu nanocomposites penetrate into eukaryotic cells, where DzFlu is internalized in the cytoplasm and predominantly in nuclei. Delivery of deoxyribozymes into cells in the proposed nanocomposites permits very efficient interactions with RNA targets inside cells. This was demonstrated by an example of inhibition of H5N1 influenza A virus replication (inhibition by a factor of ca. 3000). This effect was one order of magnitude higher than with using lipofectamine as the transfection agent. The proposed (TiO2•PL)•Dz nanocomposites demonstrated high antiviral activity and are thus potent as nucleic-acid-based drugs.

KW - Cell delivery

KW - DNAzymes

KW - Gene silencing

KW - Immobilization

KW - Influenza A virus

KW - TiO-based nanocomposites

KW - TiO2-based nanocomposites

KW - TOXICITY

KW - TITANIUM-DIOXIDE NANOPARTICLES

KW - NUCLEIC-ACIDS

KW - Humans

KW - Virus Replication/drug effects

KW - Influenza A Virus, H5N1 Subtype/drug effects

KW - DNA, Catalytic/metabolism

KW - Drug Carriers/metabolism

KW - Animals

KW - Nanocomposites

KW - Dogs

KW - Madin Darby Canine Kidney Cells

KW - HeLa Cells

KW - Antiviral Agents/metabolism

KW - Metal Nanoparticles

KW - TiO -based nanocomposites

UR - http://www.scopus.com/inward/record.url?scp=85017517124&partnerID=8YFLogxK

U2 - 10.1016/j.ijantimicag.2017.01.026

DO - 10.1016/j.ijantimicag.2017.01.026

M3 - Article

C2 - 28412273

AN - SCOPUS:85017517124

VL - 49

SP - 703

EP - 708

JO - International Journal of Antimicrobial Agents

JF - International Journal of Antimicrobial Agents

SN - 0924-8579

IS - 6

ER -