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      Development of albumin macroinitiator for polymers to use in DNA origami coating

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          Abstract

          Background/aim

          DNA nanostructures have many advantages over polymers and lipid based drug delivery agents such as biodegradability and biocompatibility. However their transfection rates and stability still limit their widely use in nanomedicine. In this study highly versatile and straightforward albumin coating preparation method is showed for DNA nanostructures.

          Materials and methods

          N-methylolmaleimide was esterified with a-bromoisobutyrl bromide (BiBB) to achive bromine functional structure. Then it was attached to bovine serum albumin (BSA) via cysteine-maleimide bond further to use as macroinitiator for atom transfer radical polymerization (ATRP). Cationic polymers can be synthesized from this end further to use as binding domain for fabricated 60 Helix bundle DNA origami.

          Results

          Proton nuclear magnetic resonance (1H NMR) analysis used for characterization. Methyelene group hydrogens’ peak in 5.0 ppm and strong peak in 1.5–2.0 ppm range showed proper methylolation of maleimide and bromine functional formation, respectively. Then BSA-macroinitiator formation is verified by 1780 Da peak shift in MALDI-TOF (Matrix-assisted laser desorption/ionization - time of flight) spectrum. Moreover electrophoretic mobility shift assay (EMSA) showed successful dense 60 Helix bundle formation.

          Conclusion

          In this study, a facile method is developed to synthesize protein conjugated-ATRP initiator further can be used in polymerization and coating DNA nanostructures. It is feasible for any protein containing cysteine amino acid.

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          Most cited references47

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          Self-assembly of DNA into nanoscale three-dimensional shapes

          Shawn M. Douglas, Hendrik Dietz, Tim Liedl … (2009)
          Molecular self-assembly offers a ‘bottom-up’ route to fabrication with subnanometre precision of complex structures from simple components1. DNA has proven a versatile building block2–5 for programmable construction of such objects, including two-dimensional crystals6, nanotubes7–11, and three-dimensional wireframe nanopolyhedra12–17. Templated self-assembly of DNA18 into custom two-dimensional shapes on the megadalton scale has been demonstrated previously with a multiple-kilobase ‘scaffold strand’ that is folded into a flat array of antiparallel helices by interactions with hundreds of oligonucleotide ‘staple strands’19, 20. Here we extend this method to building custom three-dimensional shapes formed as pleated layers of helices constrained to a honeycomb lattice. We demonstrate the design and assembly of nanostructures approximating six shapes — monolith, square nut, railed bridge, genie bottle, stacked cross, slotted cross — with precisely controlled dimensions ranging from 10 to 100 nm. We also show hierarchical assembly of structures such as homomultimeric linear tracks and of heterotrimeric wireframe icosahedra. Proper assembly requires week-long folding times and calibrated monovalent and divalent cation concentrations. We anticipate that our strategy for self-assembling custom three-dimensional shapes will provide a general route to the manufacture of sophisticated devices bearing features on the nanometer scale.
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            A logic-gated nanorobot for targeted transport of molecular payloads.

            Shawn M. Douglas, Ido Bachelet, George M. Church (2012)
            We describe an autonomous DNA nanorobot capable of transporting molecular payloads to cells, sensing cell surface inputs for conditional, triggered activation, and reconfiguring its structure for payload delivery. The device can be loaded with a variety of materials in a highly organized fashion and is controlled by an aptamer-encoded logic gate, enabling it to respond to a wide array of cues. We implemented several different logical AND gates and demonstrate their efficacy in selective regulation of nanorobot function. As a proof of principle, nanorobots loaded with combinations of antibody fragments were used in two different types of cell-signaling stimulation in tissue culture. Our prototype could inspire new designs with different selectivities and biologically active payloads for cell-targeting tasks.
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              Nucleic acid junctions and lattices.

              Nadrian Seeman (1982)
              Article 0 comments Cited 546 times – based on 0 reviews     Review now
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                Author and article information

                Journal
                Journal ID (nlm-ta): Turk J Med Sci
                Journal ID (iso-abbrev): Turk J Med Sci
                Title: Turkish Journal of Medical Sciences
                Publisher: The Scientific and Technological Research Council of Turkey
                ISSN (Print): 1300-0144
                ISSN (Electronic): 1303-6165
                Publication date Collection: 2020
                Publication date (Electronic): 26 August 2020
                Volume: 50
                Issue: 5
                Pages: 1461-1469
                Affiliations
                [1 ] Department of Nanotechnology and Nanomedicine, Graduate School of Science and Engineering, Hacettepe University, Ankara Turkey
                Author notes
                * To whom correspondence should be addressed. E-mail: saglam@ 123456hacettepe.edu.tr

                CONFLICT OF INTEREST:

                The authors declare no conflict of interest.

                Author information
                https://orcid.org/0000-0001-5288-8126
                https://orcid.org/0000-0003-3834-2260
                https://orcid.org/0000-0002-5463-8355
                Article
                DOI: 10.3906/sag-2001-245
                PMC ID: 7491299
                PubMed ID: 32283899
                SO-VID: 694ada71-f00e-4832-be90-d7de744c041f
                Copyright © Copyright © 2020 The Author(s)
                License:

                This article is distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/4.0/ ), which permits unrestricted use and redistribution provided that the original author and source are credited.

                History
                Categories
                Subject: Article

                Keywords: dna origami,bionanotechnology,atom transfer radical polymerization,drug delivery
                Data availability:
                Keywords: dna origami, bionanotechnology, atom transfer radical polymerization, drug delivery

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