Journal Home > Volume 8 , Issue 12

We previously demonstrated that polypod-like structured DNA, or polypodna, constructed with three or more oligodeoxynucleotides (ODNs), is efficiently taken up by immune cells such as dendritic cells and macrophages, depending on its structural complexity. The ODNs comprising the polypodna should bend to form the polypod-like structure, and may do so by adopting either a bendtype conformation or a cross-type conformation. Here, we tried to elucidate the orientation and bending of ODNs in polypodnas using atomic force microscopy (AFM). We designed two types of pentapodnas (i.e., a polypodna with five pods) using 60- to 88-base ODNs, which were then immobilized on DNA origami frames. AFM imaging showed that the ODNs in the pentapodna adopted bend-type conformations. Tetrapodna and hexapodna also adopted bend-type conformations when they were immobilized on frames under unconstrained conditions. These findings provide useful information toward the coherent design of, and the structure–activity relationships for, a variety of DNA nanostructures.


menu
Abstract
Full text
Outline
Electronic supplementary material
About this article

Atomic force microscopy analysis of orientation and bending of oligodeoxynucleotides in polypod-like structured DNA

Show Author's information Tomoki Shiomi1Mengmeng Tan1Natsuki Takahashi1Masayuki Endo2Tomoko Emura3Kumi Hidaka3Hiroshi Sugiyama2,3Yuki Takahashi1Yoshinobu Takakura1Makiya Nishikawa1( )
Department of Biopharmaceutics and Drug MetabolismGraduate School of Pharmaceutical Sciences, Kyoto UniversitySakyo-ku, Kyoto606-8501Japan
Institute for Integrated Cell-Material Sciences (WPI-iCeMS)Kyoto UniversitySakyo-ku, Kyoto606-8501Japan
Department of Chemistry Graduate School of Science, Kyoto UniversitySakyo-ku, Kyoto606-8502Japan

Abstract

We previously demonstrated that polypod-like structured DNA, or polypodna, constructed with three or more oligodeoxynucleotides (ODNs), is efficiently taken up by immune cells such as dendritic cells and macrophages, depending on its structural complexity. The ODNs comprising the polypodna should bend to form the polypod-like structure, and may do so by adopting either a bendtype conformation or a cross-type conformation. Here, we tried to elucidate the orientation and bending of ODNs in polypodnas using atomic force microscopy (AFM). We designed two types of pentapodnas (i.e., a polypodna with five pods) using 60- to 88-base ODNs, which were then immobilized on DNA origami frames. AFM imaging showed that the ODNs in the pentapodna adopted bend-type conformations. Tetrapodna and hexapodna also adopted bend-type conformations when they were immobilized on frames under unconstrained conditions. These findings provide useful information toward the coherent design of, and the structure–activity relationships for, a variety of DNA nanostructures.

Keywords: self-assembly, atomic force microscopy, nanotechnology, structure–activity relationship, DNA nanostructure

References(22)

1

Um, S. H.; Lee, J. B.; Park, N.; Kwon, S. Y.; Umbach, C. C.; Luo, D. Enzyme-catalysed assembly of DNA hydrogel. Nat. Mater. 2006, 5, 797–801.

2

Rothemund, P. W. K. Folding DNA to create nanoscale shapes and patterns. Nature 2006, 440, 297–302.

3

He, Y.; Ye, T.; Su, M.; Zhang, C.; Ribbe, A. E.; Jiang, W.; Mao, C. D. Hierarchical self-assembly of DNA into symmetric supramolecular polyhedra. Nature 2008, 452, 198–201.

4

Smith, D; Schüller, V; Engst, C; Rädler, J; Liedl, T. Nucleic acid nanostructures for biomedical applications. Nanomedicine 2013, 8, 105–121.

5

Um, S. H.; Lee, J. B.; Kwon, S. Y.; Li, Y. G.; Luo, D. Dendrimer-like DNA-based fluorescence nanobarcodes. Nat. Protoc. 2006, 1, 995–1000.

6

Ke, Y. G.; Ong, L. L.; Shih, W. M.; Yin, P. Three-dimensional structures self-assembled from DNA bricks. Science 2012, 338, 1177–1183.

7

Rattanakiat, S.; Nishikawa, M.; Funabashi, H.; Luo, D.; Takakura, Y. The assembly of a short linear natural cytosinephosphate-guanine DNA into dendritic structures and its effect on immunostimulatory activity. Biomaterials 2009, 30, 5701–5706.

8

Mohri, K.; Nishikawa, M.; Takahashi, N.; Shiomi, T.; Matsuoka, N.; Ogawa, K.; Endo, M.; Hidaka, K.; Sugiyama, H.; Takahashi, Y. et al. Design and development of nanosized DNA assemblies in polypod-like structures as efficient vehicles for immunostimulatory CpG motifs to immune cells. ACS Nano 2012, 6, 5931–5940.

9

Wemmer, D. E.; Wand, A. J.; Seeman, N. C.; Kallenbach, N. R. NMR analysis of DNA junctions: Imino proton NMR studies of individual arms and intact junction. Biochemistry 1985, 24, 5745–5749.

10

Ortiz-Lombardía, M.; González, A.; Eritja, R.; Aymamí, J.; Azorín, F.; Coll, M. Crystal structure of a DNA Holliday junction. Nat. Struct. Biol. 1999, 6, 913–917.

11

Lilley, D. M. Structures of helical junctions in nucleic acids. Q Rev. Biophys. 2000, 33, 109–159.

12

Li, Y. G.; Tseng, Y. D.; Kwon, S. Y.; d'Espaux, L.; Bunch, J. S.; McEuen, P. L.; Luo, D. Controlled assembly of dendrimer-like DNA. Nat. Mater. 2004, 3, 38–42.

13

Mizuno, R.; Haruta, H.; Morii, T.; Okada, T.; Asakawa, T.; Hayashi, K. Synthesis and AFM visualization of DNA nanostructures. Thin Solid Films 2004, 459–465.

14

Lyubchenko, Y. L.; Shlyakhtenko, L. S.; Ando, T. Imaging of nucleic acids with atomic force microscopy. Methods 2011, 54, 274–283.

15

Nishikawa, M.; Mizuno, Y.; Mohri, K.; Matsuoka, N.; Rattanakiat, S.; Takahashi, Y.; Funabashi, H.; Luo, D.; Takakura, Y. Biodegradable CpG DNA hydrogels for sustained delivery of doxorubicin and immunostimulatory signals in tumor-bearing mice. Biomaterials 2011, 32, 488–494.

16

Endo, M.; Katsuda, Y.; Hidaka, K.; Sugiyama, H. Regulation of DNA methylation using different tensions of double strands constructed in a defined DNA nanostructure. J. Am. Chem. Soc. 2010, 132, 1592–1597.

17

Suzuki, Y.; Endo, M.; Katsuda, Y.; Ou, K.; Hidaka, K.; Sugiyama, H. DNA origami based visualization system for studying site-specific recombination events. J. Am. Chem. Soc. 2014, 136, 211–218.

18

Murchie, A. I. H.; Clegg, R. M.; von Krtzing, E.; Duckett, D. R.; Diekmann, S.; Lilley, D. M. J. Fluorescence energy transfer shows that the four-way DNA junction is a righthanded cross of antiparallel molecules. Nature 1989, 341, 763–766.

19

Eichman, B. F.; Vargason, J. M.; Mooers, B. H. M.; Ho, P. S. The Holliday junction in an inverted repeat DNA sequence: Sequence effects on the structure of four-way junctions. Proc. Natl. Acad. Sci. USA 2000, 97, 3971–3976.

20

Sha, R.; Liu, F.; Seeman, N. C. Atomic force microscopic measurement of the interdomain angle in symmetric Holliday junctions. Biochemistry 2002, 41, 5950–5955.

21

Peters 3rd, J. P.; Maher, L. J. DNA curvature and flexibility in vitro and in vivo. Q Rev. Biophys. 2010, 43, 23–63.

22

Vafabakhsh, R.; Ha, T. Extreme bendability of DNA less than 100 base pairs long revealed by single-molecule cyclization. Science 2012, 337, 1097–1101.

File
nr-8-12-3764_ESM.pdf (1.5 MB)
Publication history
Copyright
Acknowledgements

Publication history

Received: 26 January 2015
Revised: 03 August 2015
Accepted: 06 August 2015
Published: 29 October 2015
Issue date: December 2015

Copyright

© Tsinghua University Press and Springer-Verlag Berlin Heidelberg 2015

Acknowledgements

Acknowledgements

This work was supported in part by the Grant-in-Aid for Scientific Research (B) (No. 23390010) from the Japan Society for the Promotion of Science, Grant- in-Aid for Scientific Research on Innovative Areas "Carcinogenic spiral" (No. 25114706) from the Ministry of Education, Culture, Sports, Science and Technology of Japan, and the Cross-Disciplinary Collaborative Research Promotion Project from the Institute for Integrated Cell-Material Sciences, Kyoto University.

Return