Self-assembled DNA nanostructures prepared by rolling circle amplification for the delivery of siRNA conjugates

Hong, Cheol Am; Jang, Bora; Jeong, Eun; Jeong, Hansaem; Lee, Hyukjin

doi:10.1039/c4cc03834g

Cited by 37 publications

(20 citation statements)

References 22 publications

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“…18,19,20 DNA oligomers can be used as a framework to merge several siRNA units into one construct. 21,22 DNA is especially suitable for this purpose as it is simply accessible and a huge variety of structures can be assembled by Watson Crick base pairing. As DNA is a natural molecule, it can be degraded by endogenous pathways and consequently toxicity is deemed unlikely.…”

Section: Introductionmentioning

confidence: 99%

DNA as Tunable Adaptor for siRNA Polyplex Stabilization and Functionalization

Heissig

Klein

Hadwiger³

et al. 2016

Molecular Therapy - Nucleic Acids

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siRNA and microRNA are promising therapeutic agents, which are engaged in a natural mechanism called RNA interference that modulates gene expression posttranscriptionally. For intracellular delivery of such nucleic acid triggers, we use sequence-defined cationic polymers manufactured through solid phase chemistry. They consist of an oligoethanamino amide core for siRNA complexation and optional domains for nanoparticle shielding and cell targeting. Due to the small size of siRNA, electrostatic complexes with polycations are less stable, and consequently intracellular delivery is less efficient. Here we use DNA oligomers as adaptors to increase size and charge of cargo siRNA, resulting in increased polyplex stability, which in turn boosts transfection efficiency. Extending a single siRNA with a 181-nucleotide DNA adaptor is sufficient to provide maximum gene silencing aided by cationic polymers. Interestingly, this simple strategy was far more effective than merging defined numbers (4–10) of siRNA units into one DNA scaffolded construct. For DNA attachment, the 3′ end of the siRNA passenger strand was beneficial over the 5′ end. The impact of the attachment site however was resolved by introducing bioreducible disulfides at the connection point. We also show that DNA adaptors provide the opportunity to readily link additional functional domains to siRNA. Exemplified by the covalent conjugation of the endosomolytic influenza peptide INF-7 to siRNA via a DNA backbone strand and complexing this construct with a targeting polymer, we could form a highly functional polyethylene glycol–shielded polyplex to downregulate a luciferase gene in folate receptor–positive cells.

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Section: Introductionmentioning

confidence: 99%

DNA as Tunable Adaptor for siRNA Polyplex Stabilization and Functionalization

Heissig

Klein

Hadwiger³

et al. 2016

Molecular Therapy - Nucleic Acids

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“…The 2D and 3D nanostructures can be built through complementary binding of DNA strands in a programmable manner and the simple DNA structure as “DNA brick” can grow bigger through designing complementary sticky ends . Currently, the isothermal enzymatic amplification process is a powerful tool for large‐scale preparation of DNA nanostructures, which increased the production of DNA nanostructures for more than 200 folds comparing to normal rolling circle replication with PCR . Due to the unique programmable structural design, multifunctionality and biocompatibility, DNA nanostructures have been widely applied in biosensing, bioimaging, drug delivery, molecular computation, and as macromolecular scaffold .…”

Section: Biomedical Applicationsmentioning

confidence: 99%

Programmable and Multifunctional DNA‐Based Materials for Biomedical Applications

et al. 2018

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DNA encodes the genetic information; recently, it has also become a key player in material science. Given the specific Watson-Crick base-pairing interactions between only four types of nucleotides, well-designed DNA self-assembly can be programmable and predictable. Stem-loops, sticky ends, Holliday junctions, DNA tiles, and lattices are typical motifs for forming DNA-based structures. The oligonucleotides experience thermal annealing in a near-neutral buffer containing a divalent cation (usually Mg ) to produce a variety of DNA nanostructures. These structures not only show beautiful landscape, but can also be endowed with multifaceted functionalities. This Review begins with the fundamental characterization and evolutionary trajectory of DNA-based artificial structures, but concentrates on their biomedical applications. The coverage spans from controlled drug delivery to high therapeutic profile and accurate diagnosis. A variety of DNA-based materials, including aptamers, hydrogels, origamis, and tetrahedrons, are widely utilized in different biomedical fields. In addition, to achieve better performance and functionality, material hybridization is widely witnessed, and DNA nanostructure modification is also discussed. Although there are impressive advances and high expectations, the development of DNA-based structures/technologies is still hindered by several commonly recognized challenges, such as nuclease instability, lack of pharmacokinetics data, and relatively high synthesis cost.

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“…Because only the scaffold is synthesized by RCA, separate preparation of a set of staple strands is required. A four‐arm motif and a Y‐shaped DNA motif have also been fabricated by RCA. In these studies, the long DNA strand produced was cleaved by a restriction enzyme to produce structural motifs.…”

Section: Introductionmentioning

confidence: 99%

Construction of T‐Motif‐Based DNA Nanostructures through Enzymatic Reactions

et al. 2018

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The most common way to fabricate DNA nanostructures is to mix individually synthesized DNA oligomers in one pot. However, if DNA nanostructures could be produced through enzymatic reactions, they could be applied in various environments, including in vivo. Herein, an enzymatic method developed to construct a DNA nanostructure from a simple motif called a T-motif is reported. A long, repeated structure was replicated from a circular template by rolling circle amplification and then cleaved into T-motif segments by restriction enzymes. These motifs have been successfully assembled into a ladder-like nanostructure without purification or controlled annealing. This approach is widely applicable to constructing a variety of DNA nanostructures through enzymatic reactions.

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Self-assembled DNA nanostructures prepared by rolling circle amplification for the delivery of siRNA conjugates

Cited by 37 publications

References 22 publications

DNA as Tunable Adaptor for siRNA Polyplex Stabilization and Functionalization

DNA as Tunable Adaptor for siRNA Polyplex Stabilization and Functionalization

Programmable and Multifunctional DNA‐Based Materials for Biomedical Applications

Construction of T‐Motif‐Based DNA Nanostructures through Enzymatic Reactions

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