A practical approach for the detection of DNA nanostructures in single live human cells by fluorescence microscopy.
In the last decade, in vivo studies have revealed that even subtle differences in size, concentration of components, cell cycle stage, make the cells in a population respond differently to the same stimulus. In order to characterize such complexity of behavior and shed more light on the functioning and communication amongst cells, researchers are developing strategies to study single live cells in a population. In this paper, we describe the methods to design and prepare DNA-based fluorescent tetrahedral nanostructures, to deliver them to live cells and characterize such cells with epifluorescence microscopy. We report that HeLa cells internalize these nanostructures spontaneously with a higher efficiency with respect to single-stranded or double-stranded oligonucleotides. Our findings suggest that DNA tetrahedra could serve as a platform for the realization of a series of multifunctional intracellular biosensors for the analysis of single live cells.
Interdepartmental Center for Industrial Research, Health Sciences & Technologies (CIRI-HST) at the University of Bologna, Via Tolara di Sopra 41/E, Ozzano dell'Emilia, Bologna 40064, Italy.
C
C
Bergamini
Department of Pharmacy and Biotechnologies (FaBiT), University of Bologna, Italy.
P
P
Angelini
Interdepartmental Center for Industrial Research, Health Sciences & Technologies (CIRI-HST) at the University of Bologna, Via Tolara di Sopra 41/E, Ozzano dell'Emilia, Bologna 40064, Italy; Department of Medical and Surgical Sciences, University of Bologna, Italy.
K J
KJ
Rhoden
Interdepartmental Center for Industrial Research, Health Sciences & Technologies (CIRI-HST) at the University of Bologna, Via Tolara di Sopra 41/E, Ozzano dell'Emilia, Bologna 40064, Italy; Department of Pharmacy and Biotechnologies (FaBiT), University of Bologna, Italy.
A M
AM
Porcelli
Interdepartmental Center for Industrial Research, Health Sciences & Technologies (CIRI-HST) at the University of Bologna, Via Tolara di Sopra 41/E, Ozzano dell'Emilia, Bologna 40064, Italy; Department of Pharmacy and Biotechnologies (FaBiT), University of Bologna, Italy.
R
R
Fato
Interdepartmental Center for Industrial Research, Health Sciences & Technologies (CIRI-HST) at the University of Bologna, Via Tolara di Sopra 41/E, Ozzano dell'Emilia, Bologna 40064, Italy; Department of Pharmacy and Biotechnologies (FaBiT), University of Bologna, Italy; Italian National Research Council (CNR), Istituto Nanoscienze, S3 Center, Modena, Italy; National Interuniversity Consortium of Materials Science and Technology (INSTM) at the University of Bologna, Italy. Electronic address: giampaolo.zuccheri@unibo.it.
G
G
Zuccheri
D004247:DNA / Q000737:chemistry* / Q000648:ultrastructure; D005456:Fluorescent Dyes / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D008856:Microscopy, Fluorescence; D049329:Nanostructures / Q000737:chemistry / Q000648:ultrastructure; D009690:Nucleic Acid Conformation; D009841:Oligonucleotides / Q000737:chemistry*
D005456:Fluorescent Dyes; D009841:Oligonucleotides; D004247:DNA
DNA nanotechnology; Flow cytofluorimetry; Fluorescence microscopy; Live-cell imaging; Self-assembled nanostructures