Direct visualization of the drug loading of single DNA origami nanostructures by AFM-IR nanospectroscopy

Hanke, Marcel; Grundmeier, Guido; Keller, Adrian

doi:10.1039/d2nr02701a

Cited by 9 publications

(10 citation statements)

References 51 publications

(88 reference statements)

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“…Despite these limitations, the enhanced contrast and observed contribution of Pt in the EDS spectra of cisplatin-loaded DONs (when compared to pure DON) already demonstrated cisplatin binding. Recent attempts to probe drug loading onto DONs using AFM-IR nanospectroscopy can be a promising alternative method, although it is limited to drug molecules with strong IR signals and for the moment, does not offer enough spatial resolution to map binding sites …”

Section: Resultsmentioning

confidence: 99%

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Cisplatin-Cross-Linked DNA Origami Nanostructures for Drug Delivery Applications

Sala

Perečko

Mestek

et al. 2022

ACS Appl. Nano Mater.

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Biocompatible DNA origami nanostructures (DONs) attract significant interest as potential targeted drug carriers. Their sensitivity to various environmental triggers is beneficial as a drug release or decomposition mechanism, but it also hinders their stability; therefore, they require some extent of cross-linking. Using cisplatin, we demonstrate that a molecule can both act as a therapeutic and cross-linking agent for DONs. In this work, triangular two-dimensional DONs are loaded with ∼1000 cisplatin molecules per nanostructure as confirmed by ICP-MS and STEM-EDS. Time dependence of the loading shows a saturation point and gradual cisplatin release. Above ∼1000 cisplatin molecules loaded per nanostructure, structural distortion occurs as analyzed by AFM and gel electrophoresis. Cross-linking persists even after thermal treatment of cisplatin-loaded DONs above typical DON denaturing temperatures preventing complete separation into DON components (scaffold and staples). Cisplatin-loaded DON cytotoxicity is tested on FaDu cells and compared to that of free cisplatin using MTT assays. Nanomolar quantities of cisplatin-loaded DONs reduce cell viability to 50% after 48–72 h. Considering the low dose required of cisplatin-loaded DONs to exhibit cytotoxic behavior and the known tumor-targeting properties of DONs, this system can be a promising drug carrier for cancer therapy worthy of further exploration.

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Section: Resultsmentioning

confidence: 99%

“…Recent attempts to probe drug loading onto DONs using AFM-IR nanospectroscopy can be a promising alternative method, although it is limited to drug molecules with strong IR signals and for the moment, does not offer enough spatial resolution to map binding sites. 48 3.5. Thermal Stability of Cisplatin-Loaded DONs.…”

Section: Elemental Map Of Cisplatin-loaded Donsmentioning

confidence: 99%

Cisplatin-Cross-Linked DNA Origami Nanostructures for Drug Delivery Applications

Sala

Perečko

Mestek

et al. 2022

ACS Appl. Nano Mater.

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“…DNA origami assembly : The DNA origami nanostructures (“tall” rectangle, [1] Rothemund triangle, [1] 6HB 42‐bpCS, [14] 6HB 21‐bpCS, [14] 24HB) [24] were assembled as previously described [36] in 1xTAE buffer (Carl Roth) containing 10 mM MgCl 2 (Sigma‐Aldrich) at tenfold excess of staples (Eurofins) to scaffold (Tilibit). In the assembly of the rectangle, all edge staples were omitted in order to avoid aggregation due to blunt‐end stacking [1] .…”

Section: Methodsmentioning

confidence: 99%

Effect of Ionic Strength on the Thermal Stability of DNA Origami Nanostructures

et al. 2023

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The stability of DNA origami nanostructures in aqueous media is closely tied to the presence of cations that screen electrostatic inter‐helix repulsion. Here, the thermal melting behavior of different DNA origami nanostructures is investigated in dependence on Mg2+ concentration and compared to calculated ensemble melting temperatures of the staple strands used in DNA origami folding. Strong deviations of the measured DNA origami melting temperatures from the calculated ones are observed, in particular at high ionic strength where the melting temperature saturates and becomes independent of ionic strength. The degree of deviation between the measured and calculated melting temperatures further depends on the superstructure and in particular the mechanical properties of the DNA origami nanostructures. This indicates that thermal stability of a given DNA origami design at high ionic strength is governed predominantly not by electrostatic inter‐helix repulsion but mostly by mechanical strain.

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“…For example, UV absorbance rapidly provides a measure of concentration under the assumption of a well-formed structure. 39,40 Gel electrophoresis is a low cost but slow assay that can distinguish between different morphologies of nanostructures with the same molecular weight. Gel band intensity can also be related to yield or concentration, while the spread or smearing of a band can speak with formation and degradation.…”

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Capacitance measurements for assessing DNA origami nanostructures

Walawalkar

Sajal

Gilpin

et al. 2023

Preprint

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Nanostructures fabricated with DNA are emerging as a practical approach for applications ranging from advanced manufacturing to therapeutics. To support the strides made in improving accessibility and facilitating commercialization of DNA nanostructure applications, we identify the need for a rapid characterization approach that aids nanostructure production. In our work, we introduce a low-fidelity characterization approach that provides an interdependent assessment of DNA origami formation, concentration and morphology using capacitance sensing. Change in charge is one of the transduction methods to determine capacitive loading on a substrate. It is known that cations in the solution stabilize DNA origami nanostructures. So, we hypothesized that the presence of cations and nanostructures in a buffer solution can induce capacitance change that is distinctive of the nanostructure present. In this study we were able to detect a change in the capacitance when the nanostructure solution was deposited on our capacitance sensor, and we could distinguish between pre-annealed and annealed structures at concentrations less than 15 nM. The capacitance measurements were affected by the concentration of Mg2+ions in the solution, the staple-to-scaffold stoichiometric ratio of the nanostructure and the nanostructure morphology. Maintaining a 12.5 mM Mg2+concentration in the nanostructure buffer, we discover a linear relationship between the relative capacitance change and the nanostructure concentration from 5 nM to 20 nM, which we call the characteristic curve. We find distinct characteristic curves for our three nanostructures with distinct morphologies but similar molecular weight - a rectangular plate, a sphere and a rod. Given that we can distinguish nanostructure formation, concentration and morphology, we expect that capacitance measurement will emerge as an affordable and rapid approach for quality control for nanostructure production.

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Direct visualization of the drug loading of single DNA origami nanostructures by AFM-IR nanospectroscopy

Abstract: The efficient loading of DNA nanostructures with intercalating or groove-binding drugs is an important prerequisite for various applications in drug delivery. However, unambiguous verification and quantification of successful drug loading...

Cited by 9 publications

References 51 publications

Cisplatin-Cross-Linked DNA Origami Nanostructures for Drug Delivery Applications

Cisplatin-Cross-Linked DNA Origami Nanostructures for Drug Delivery Applications

Effect of Ionic Strength on the Thermal Stability of DNA Origami Nanostructures

Capacitance measurements for assessing DNA origami nanostructures

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