title: Folding and Characterization of a Bio-responsive Robot from DNA Origami.
abstract: The DNA nanorobot is a hollow hexagonal nanometric device, designed to open in response to specific stimuli and present cargo sequestered inside. Both stimuli and cargo can be tailored according to specific needs. Here we describe the DNA nanorobot fabrication protocol, with the use of the DNA origami technique. The procedure initiates by mixing short single-strand DNA staples into a stock mixture which is then added to a long, circular, single-strand DNA scaffold in presence of a folding buffer. A standard thermo cycler is programmed to gradually lower the mixing reaction temperature to facilitate the staples-to-scaffold annealing, which is the guiding force behind the folding of the nanorobot. Once the 60 hr folding reaction is complete, excess staples are discarded using a centrifugal filter, followed by visualization via agarose-gel electrophoresis (AGE). Finally, successful fabrication of the nanorobot is verified by transmission electron microscopy (TEM), with the use of uranyl-formate as negative stain.
authors.affiliation: Faculty of Life Sciences and the Institute for Nanotechnology & Advanced Materials, Bar-Ilan University.
authors.forename: Yaniv
authors.initials: Y
authors.lastname: Amir
authors.affiliation: Faculty of Life Sciences and the Institute for Nanotechnology & Advanced Materials, Bar-Ilan University.
authors.forename: Almogit
authors.initials: A
authors.lastname: Abu-Horowitz
authors.affiliation: Faculty of Life Sciences and the Institute for Nanotechnology & Advanced Materials, Bar-Ilan University; ido.bachelet@biu.ac.il.
authors.forename: Ido
authors.initials: I
authors.lastname: Bachelet
mesh_terms: D004247:DNA / Q000737:chemistry*; D004277:DNA, Single-Stranded / Q000737:chemistry; D004587:Electrophoresis, Agar Gel; D046529:Microscopy, Electron, Transmission; D049329:Nanostructures / Q000737:chemistry; D009690:Nucleic Acid Conformation; D012371:Robotics; D013194:Staining and Labeling / Q000379:methods
chemical_list: D004277:DNA, Single-Stranded; D004247:DNA