title: Imaging of transfection and intracellular release of intact, non-labeled DNA using fluorescent nanodiamonds.
abstract: 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 biosensors reporting complex molecular events.
authors.affiliation: Faculty of Biomedical Engineering, Czech Technical University in Prague, Sitna sq. 3105, 272 01 Kladno, Czech Republic and Institute of Physics AS CR, v.v.i, Na Slovance 1999/2, 182 21 Prague 8, Czech Republic.
authors.forename: V
authors.initials: V
authors.lastname: Petrakova
authors.affiliation: Faculty of Biomedical Engineering, Czech Technical University in Prague, Sitna sq. 3105, 272 01 Kladno, Czech Republic and Institute of Microbiology AS CR, v.v.i, Videnska 1083, 142 20 Prague 4, Czech Republic.
authors.forename: V
authors.initials: V
authors.lastname: Benson
authors.affiliation: Generi Biotech Ltd., Machkova 587, 500 11 Hradec Kralove, Czech Republic.
authors.forename: M
authors.initials: M
authors.lastname: Buncek
authors.affiliation: Faculty of Biomedical Engineering, Czech Technical University in Prague, Sitna sq. 3105, 272 01 Kladno, Czech Republic and Institute of Microbiology AS CR, v.v.i, Videnska 1083, 142 20 Prague 4, Czech Republic.
authors.forename: A
authors.initials: A
authors.lastname: Fiserova
authors.affiliation: Faculty of Biomedical Engineering, Czech Technical University in Prague, Sitna sq. 3105, 272 01 Kladno, Czech Republic and Institute of Organic Chemistry and Biochemistry AS CR, v.v.i., Flemingovo nam. 2, 166 10 Prague 6, Czech Republic. cigler@uochb.cas.cz.
authors.forename: M
authors.initials: M
authors.lastname: Ledvina
authors.affiliation: Nuclear Physics Institute AS CR, v.v.i., 250 68, Rez near Prague, Czech Republic.
authors.forename: J
authors.initials: J
authors.lastname: Stursa
authors.affiliation: Institute of Organic Chemistry and Biochemistry AS CR, v.v.i., Flemingovo nam. 2, 166 10 Prague 6, Czech Republic. cigler@uochb.cas.cz.
authors.forename: P
authors.initials: P
authors.lastname: Cigler
authors.affiliation: Faculty of Biomedical Engineering, Czech Technical University in Prague, Sitna sq. 3105, 272 01 Kladno, Czech Republic and IMEC Division IMOMEC, Hasselt University, Wetenschapspark 1, B-3590, Diepenbeek, Belgium and Institute for Materials Research, Hasselt University, Wetenschapspark 1, B-3590 Diepenbeek, Belgium. milos.nesladek@uhasselt.be.
authors.forename: M
authors.initials: M
authors.lastname: Nesladek
mesh_terms: D000818:Animals; D004247:DNA; D019073:HT29 Cells; D006801:Humans; D049449:Luminescence; D008811:Mice, Inbred DBA; D058612:Nanodiamonds; D014162:Transfection
chemical_list: D058612:Nanodiamonds; D004247:DNA