Univerzita Pardubice Fakulta chemicko- technologická

Univerzita Pardubice

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Publikace detail

Micro-RNA isolation using TiO2 materials

Autoři: Smělá Denisa | Kupčík Rudolf | Zelinková Eliška | Macák Jan | Bílková Zuzana

Rok: 2019

Druh publikace: ostatní - přednáška nebo poster

Strana od-do: nestránkováno

Tituly:

Jazyk	Název	Abstrakt	Klíčová slova
eng	Micro-RNA isolation using TiO2 materials	Micro-RNAs (miRNAs) are endogenous, short (~23 nucleotides), non-coding molecules of RNA. They partake in regulation of gene expression and various biological processes. Expression of miRNAs differs for healthy and pathological conditions. Therefore, miRNAs are considered very promising diagnostic or prognostic biomarkers for several diseases, i.e. carcinogenesis. Isolation and detection of miRNAs are difficult due to their small sizes, low concentrations (in fM) and high homology in sequences. Commonly used methods for miRNA isolation are phenol-chloroform extraction and solid-phase extraction. It was described, that materials based on TiO2 have an affinity to nucleic acids thanks to their strong interactions with phosphate backbone. Nucleic acids are negatively charged and charge of TiO2 under acidic conditions is strong positive. In addition, it was suggested, that the adsorption of DNA by TiO2 is caused by interaction between DNA and hydroxyl groups on the surface of TiO2. Similar principles apply for TiO2 interactions with RNA. In our work, we tested two different materials based on TiO2 to determine their applicability for miRNA isolation - 10 µm TiO2 microspheres (Titansphere (TiO) - GL Sciences) and TiO2 nanotubes coated with Fe3O4 (TiO2NTs@Fe3O4NPs, CEMNAT). Those were tested on the mixture of genomic RNA pre-isolated from Jurkat cells (T-lymphoblast cell line) spiked with oligo RNA standard based on hsa-miR-18a-3p (Generi Biotech). We optimized binding and elution conditions for extraction of short RNAs from this mixture containing wide range of RNA sizes.	miRNA; solid-phase extraction; titanium dioxide nanomaterials