Minimizing Structural Heterogeneity in DNA Self-Assembled Dye Templating via DNA Origami-Tuned Conformations.

With recent advances in DNA-templated dye aggregation for leveraging and engineering molecular excitons, a need exists for minimizing structural heterogeneity. Holliday Junction complexes (HJ) are commonly used to covalently template dye aggregates on their core; however, the global conformation of HJ is detrimentally dynamic. Here, the global conformation of the HJ is selectively tuned by restricting its position and orientation by using a sheet-like DNA origami construct (DOC) physisorbed on glass. The HJ arms are fixed with four different designed interduplex angles (IDAs). Atomic force microscopy confirmed that the HJs are bound to the surface of DOC with tuned IDAs. Dye orientation distributions were determined by combining dipole imaging and super-resolution microscopy. All IDAs led to dye orientations having dispersed distributions along planes perpendicular to the HJ plane, suggesting that stacking occurred between the dye and the neighboring DNA bases. The dye-base stacking interpretation was supported by increasing the size of the core cavity. The narrowest IDA minimizes structural heterogeneity and suggests dye intercalation. A strong correlation is found between the IDA and the orientation of the dye along the HJ plane. These results show that the HJ imposes restrictions on the dye and that the dye-DNA interactions are always present regardless of global conformation. The implications of our results are discussed for the scalability of dye aggregates using DNA self-assembly. Our methodology provides an avenue for the solid-supported single-molecule characterization of molecular assemblies templated on biomolecules─such as DNA and protein templates involved in light-harvesting and catalysis─with tuned conformations and restricted in position and orientation.

Effect of hydrophilicity-imparting substituents on exciton delocalization in squaraine dye aggregates covalently templated to DNA Holliday junctions.

Molecular aggregates exhibit emergent properties, including the collective sharing of electronic excitation energy known as exciton delocalization, that can be leveraged in applications such as quantum computing, optical information processing, and light harvesting. In a previous study, we found unexpectedly large excitonic interactions (quantified by the excitonic hopping parameter Jm,n) in DNA-templated aggregates of squaraine (SQ) dyes with hydrophilic-imparting sulfo and butylsulfo substituents. Here, we characterize DNA Holliday junction (DNA-HJ) templated aggregates of an expanded set of SQs and evaluate their optical properties in the context of structural heterogeneity. Specifically, we characterized the orientation of and Jm,n between dyes in dimer aggregates of non-chlorinated and chlorinated SQs. Three new chlorinated SQs that feature a varying number of butylsulfo substituents were synthesized and attached to a DNA-HJ via a covalent linker to form adjacent and transverse dimers. Various characteristics of the dye, including its hydrophilicity (in terms of log Po/w) and surface area, and of the substituents, including their local bulkiness and electron withdrawing capacity, were quantified computationally. The orientation of and Jm,n between the dyes were estimated using a model based on Kühn-Renger-May theory to fit the absorption and circular dichroism spectra. The results suggested that adjacent dimer aggregates of all the non-chlorinated and of the most hydrophilic chlorinated SQ dyes exhibit heterogeneity; that is, they form a mixture of dimers subpopulations. A key finding of this work is that dyes with a higher hydrophilicity (lower log Po/w) formed dimers with smaller Jm,n and large center-to-center dye distance (Rm,n). Also, the results revealed that the position of the dye in the DNA-HJ template, that is, adjacent or transverse, impacted Jm,n. Lastly, we found that Jm,n between symmetrically substituted dyes was reduced by increasing the local bulkiness of the substituent. This work provides insights into how to maintain strong excitonic coupling and identifies challenges associated with heterogeneity, which will help to improve control of these dye aggregates and move forward their potential application as quantum information systems.

Towards control of excitonic coupling in DNA-templated Cy5 aggregates: the principal role of chemical substituent hydrophobicity and steric interactions.

Understanding and controlling exciton coupling in dye aggregates has become a greater focus as potential applications such as coherent exciton devices, nanophotonics, and biosensing have been proposed. DNA nanostructure templates allow for a powerful modular approach. Using DNA Holliday junction (HJ) templates variations of dye combinations and precision dye positions can be rapidly assayed, as well as creating aggregates of dyes that could not be prepared (either due to excess or lack of solubility) through alternative means. Indodicarbocyanines (Cy5) have been studied in coupled systems due to their large transition dipole moment, which contributes to strong coupling. Cy5-R dyes were recently prepared by chemically modifying the 5,5'-substituents of indole rings, resulting in varying dye hydrophobicity/hydrophilicity, steric considerations, and electron-donating/withdrawing character. We utilized Cy5-R dyes to examine the formation and properties of 30 unique DNA templated homodimers. We find that in our system the sterics of Cy5-R dyes play the determining factor in orientation and coupling strength of dimers, with coupling strengths ranging from 50-138 meV. The hydrophobic properties of the Cy5-R modify the percentage of dimers formed, and have a secondary role in determining the packing characteristics of the dimers when sterics are equivalent. Similar to other reports, we find that positioning of the Cy5-R within the HJ template can favor particular dimer interactions, specifically oblique or H-type dimers.

The water temperature changes the effect of pH on copper toxicity to the green microalgae Raphidocelis subcapitata.

Rising temperature enhances the algal growth, which in turn increases the water pH. Ecotoxicity studies have suggested that copper becomes more toxic to microalgae species by increasing the temperature (within 20-30 °C) and pH. In this study, the joined effect of pH and temperature on copper toxicity to the microalgae Raphidocelis subcapitata was investigated using acclimated cells. Algal growth and toxicity tests were conducted using the medium recommended by the Organisation for Economic Co-operation and Development (OECD medium) at pH 6, 7, and 8 units from 15 to 30 °C, spaced by 3 °C. The specific growth rate of R. subcapitata increased by raising the pH and temperature, attributed to the higher membrane permeability and metabolism. The ecotoxicity tests showed that temperature changes the effect of pH on copper toxicity. Copper became less toxic when rising the temperature from 15 to 18 °C and from 6 to 8 pH-unit, suggesting that high pH controls copper bioavailability and toxicity. In contrast, from 21 to 30 °C, the effect of copper was not significantly altered by temperature, but it became more toxic at high pH. Results of this study warn about the higher risk of copper in cold seasons rather than warm conditions.

Effects of chemical interaction of nutrients and EDTA on metals toxicity to Pseudokirckneriella subcapitata.

We studied the effect of the chemical interaction of nutrients and the ethylenediamine tetraacetic acid (EDTA) on metals toxicity. Growth inhibition tests of Pseudokirchneriella subcapitata by nutrient metals copper (Cu) and zinc (Zn), and the non-nutrient metal lead (Pb), were performed. The high-enriched Bold's Basal medium (BBm) and two low-enriched standard media, recommended by the Organization for Economic Cooperation and Development (OECDm) and Environmental Protection Agency-algal assay procedure medium (AAPm), were used in this study. The metals toxicity was affected by the interaction of nutrients and EDTA. Cu+2 was more toxic in the OECDm (EC50 20.3 µg/L), while Pb+2 (EC50 23.1 µg/L) and Zn+2 (EC50 99.4 µg/L) in the AAPm. Non-toxic effect of these metals was observed in BBm, but the exclusion of EDTA shifted it into a toxic medium. Finally, we found that the toxicity of the studied nutrient metals is mainly influenced by EDTA, which reduced the concentration of ionized metals, while the toxicity of the non-nutrient metal is affected by EDTA and phosphates.