title: Precise Coating of a Wide Range of DNA Templates by a Protein Polymer with a DNA Binding Domain.
abstract: Emerging DNA-based nanotechnologies would benefit from the ability to modulate the properties (e.g., solubility, melting temperature, chemical stability) of diverse DNA templates (single molecules or origami nanostructures) through controlled, self-assembling coatings. We here introduce a DNA coating agent, called C8-BSso7d, which binds to and coats with high specificity and affinity, individual DNA molecules as well as folded origami nanostructures. C8-BSso7dcoats and protects without condensing, collapsing or destroying the spatial structure of the underlying DNA template. C8-BSso7dcombines the specific nonelectrostatic DNA binding affinity of an archeal-derived DNA binding domain (Sso7d, 7 kDa) with a long hydrophilic random coil polypeptide (C8, 73 kDa), which provides colloidal stability (solubility) through formation of polymer brushes around the DNA templates. C8-BSso7dis produced recombinantly in yeast and has a precise (but engineerable) amino acid sequence of precise length. Using electrophoresis, AFM, and fluorescence microscopy we demonstrate protein coat formation with stiffening of one-dimensional templates (linear dsDNA, supercoiled dsDNA and circular ssDNA), as well as coat formation without any structural distortion or disruption of two-dimensional DNA origami template. Combining the programmability of DNA with the nonperturbing precise coating capability of the engineered protein C8-BSso7dholds promise for future applications such as the creation of DNA-protein hybrid networks, or the efficient transfection of individual DNA nanostructures into cells.
authors.affiliation: Physical Chemistry and Soft Matter, Wageningen University and Research , Stippeneng 4, 6708 WE Wageningen, The Netherlands.
authors.forename: Armando
authors.identifier: 0000-0001-5505-3423
authors.initials: A
authors.lastname: Hernandez-Garcia
authors.affiliation: Department of Materials Science and Engineering, North Carolina State University , Raleigh, North Carolina 27695, United States.
authors.forename: Nicole A
authors.initials: NA
authors.lastname: Estrich
authors.affiliation: Wageningen UR Food and Biobased Research, Wageningen University and Research , Bornse Weilanden 9, 6708 WG, Wageningen, The Netherlands.
authors.forename: Marc W T
authors.initials: MW
authors.lastname: Werten
authors.affiliation: Department of Physics, National University of Singapore , 2 Science Drive 3, 117542 Singapore.
authors.forename: Johan R C
authors.initials: JR
authors.lastname: Van Der Maarel
authors.affiliation: Department of Materials Science and Engineering, North Carolina State University , Raleigh, North Carolina 27695, United States.
authors.forename: Thomas H
authors.identifier: 0000-0002-6739-2059
authors.initials: TH
authors.lastname: LaBean
authors.affiliation: Wageningen UR Food and Biobased Research, Wageningen University and Research , Bornse Weilanden 9, 6708 WG, Wageningen, The Netherlands.
authors.forename: Frits A
authors.initials: FA
authors.lastname: de Wolf
authors.affiliation: Physical Chemistry and Soft Matter, Wageningen University and Research , Stippeneng 4, 6708 WE Wageningen, The Netherlands.
authors.forename: Martien A
authors.initials: MA
authors.lastname: Cohen Stuart
authors.affiliation: Physical Chemistry and Soft Matter, Wageningen University and Research , Stippeneng 4, 6708 WE Wageningen, The Netherlands.
authors.forename: Renko
authors.initials: R
authors.lastname: de Vries
mesh_terms: D001665:Binding Sites; D004247:DNA / Q000737:chemistry*; D008958:Models, Molecular; D049329:Nanostructures / Q000737:chemistry; D036103:Nanotechnology; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry*; D011487:Protein Conformation; D011506:Proteins / Q000138:chemical synthesis / Q000737:chemistry*
chemical_list: D010455:Peptides; D011506:Proteins; D004247:DNA
keywords: DNA nanotechnology; directed self-assembly; nanomaterials; protein engineering; protein polymer; single molecule