Self‐Assembly of DNA Origami Heterodimers in High Yields and Analysis of the Involved Mechanisms. Issue 51 (21st November 2019)

• Record Type: Journal Article
• Authors: Sheheade, Breveruos; Liber, Miran; Popov, Mary; Berger, Yaron; Khara, Dinesh C.; Jopp, Jürgen; Nir, Eyal
• Is Part Of: Small. Volume 15:Issue 51(2019)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2019-11-21
• Subjects: agarose gel electrophoresis -- DNA molecular devices -- DNA nanotechnology -- DNA origami; Nanotechnology -- Periodicals; Nanoparticles -- Periodicals; Microtechnology -- Periodicals; 620.5
• DOI: 10.1002/smll.201902979
• Languages: English
• ISSNs: 1613-6810

Abstract

Abstract: Efficient fabrication of structurally and functionally diverse nanomolecular devices and machines by organizing separately prepared DNA origami building blocks into a larger structure is limited by origami attachment yields. A general method that enables attachment of origami building blocks using 'sticky ends' at very high yields is demonstrated. Two different rectangular origami monomers are purified using agarose gel electrophoresis conducted in solute containing 100 × 10 −3 m NaCl, a treatment that facilitates the dissociation of most of the incorrectly hybridized origami structures that form through blunt‐end interactions during the thermal annealing process and removes these structures as well as excess strands that otherwise interfere with the desired heterodimerization reaction. Heterodimerization yields of gel‐purified monomers are between 98.6% and 99.6%, considerably higher than that of monomers purified using the polyethylene glycol (PEG) method (88.7–96.7%). Depending on the number of PEG purification rounds, these results correspond to about 4‐ to 25‐fold reduction in the number of incorrect structures observed by atomic force microscopy. Furthermore, the analyses of the incorrect structures observed before and after the heterodimerization reactions and comparison of the purification methods provide valuable information on the reaction mechanisms that interfere with heterodimerization. Abstract : DNA origami dimerization yield is typically not very high. Purification of rectangle origami monomers using agarose gel electrophoresis reduces unreactive monomers and incorrect structures by 4‐ to 25‐fold compared to polyethylene‐glycol‐purified samples, achieving 99.6% heterodimerization yields.