Imaging of transfection and intracellular release of intact, non-labeled DNA using fluorescent nanodiamonds
| Autor: | Petr Cigler, Milos Nesladek, Veronika Benson, Martin Bunček, A. Fiserova, M. Ledvina, Vladimira Petrakova, Jan Stursa |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
Luminescence
Materials science Ethyleneimine Nanotechnology DNA 02 engineering and technology Transfection 010402 general chemistry 021001 nanoscience & nanotechnology 01 natural sciences Fluorescence Nanodiamonds 0104 chemical sciences Mice Inbred DBA Animals Humans General Materials Science 0210 nano-technology Nanodiamond HT29 Cells Nanoscopic scale Biosensor Intracellular |
| Popis: | Efficient delivery of stabilized nucleic acids (NAs) into cells and release of the NA payload are crucial points in the transfection process. Here we report on the fabrication of a nanoscopic cellular delivery carrier that is additionally combined with a label-free intracellular sensor device, based on biocompatible fluorescent nanodiamond particles. The sensing function is engineered into nanodiamonds by using nitrogen-vacancy color centers, providing stable non-blinking luminescence. The device is used for monitoring NA transfection and the payload release in cells. The unpacking of NAs from a poly(ethyleneimine)-terminated nanodiamond surface is monitored using the color shift of nitrogen-vacancy centers in the diamond, which serve as a nanoscopic electric charge sensor. The proposed device innovates the strategies for NA imaging and delivery, by providing detection of the intracellular release of non-labeled NAs without affecting cellular processing of the NAs. Our system highlights the potential of nanodiamonds to act not merely as labels but also as non-toxic and non-photobleachable fluorescent bio-sensors reporting complex molecular events. This work was supported by Ministry of Health of the Czech Republic, grant no. 15e33094A and ESF European social fund in the Czech Republic - Investments in Education Development - Grant No. CZ.1.07/2.3.00/20.0306. The work of M. N. was supported by the FWO project (Flanders Diamond spin-magnetometers and nano-particle FRET sensors). Irradiation of NDs was carried out at the CANAM infrastructure of the NPI ASCR Rez supported through MSMT project no. LM2011019. We thank Dr J. Micova, Dr I. Rehor, Mr J. Havlik and Mr M. Gulka for their very valuable help and Dr H. Hoffman for critical proofreading of the manuscript. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|