Formulation of DNA Nanocomposites: Towards Functional Materials for Protein Expression

Abstract

DNA hydrogels are an emerging class of materials that hold great promise for numerous biotechnological applications, ranging from tissue engineering to targeted drug delivery and cell-free protein synthesis (CFPS). In addition to the molecular programmability of DNA that can be used to instruct biological systems, the formulation of DNA materials, e.g., as bulk hydrogels or microgels, is also relevant for specific applications. To advance the state of knowledge in this research area, the present work explores the scope of a recently developed class of complex DNA nanocomposites, synthesized by RCA polymerization of DNA-functionalized silica nanoparticles (SiNPs) and carbon nanotubes (CNTs). SiNP/CNT–DNA composites were produced as bulk materials and microgels which contained a plasmid with transcribable genetic information for a fluorescent marker protein. Using confocal microscopy and flow cytometry, we found that the materials are very efficiently taken up by various eukaryotic cell lines, which were able to continue dividing while the ingested material was evenly distributed to the daughter cells. However, no expression of the encoded protein occurred within the cells. While the microgels did not induce production of the marker protein even in a CFPS procedure with eukaryotic cell lysate, the bulk composites proved to be efficient templates for CFPS. This work contributes to the understanding of the molecular interactions between DNA composites and the functional cellular machinery. Implications for the use of such materials for CFPS procedures are discussed.

Macrophage-Targeting DNA Nanomaterials: A Future Direction of Biological Therapy

“…53 Schipperkes et al developed multifunctional DNA nanomaterials consisting of silica nanoparticles and carbon nanotubes functionalized with DNA. 54 Because their characteristics can be programmed and modified, DNA nanomaterials can be used as substance carriers for therapeutic purposes. DNA nanomaterials loaded with pharmacological compounds, ribonucleic acids, and antibiotics show high load capacities and can have different effects.…”

Exploring the diverse biomedical applications of programmable and multifunctional DNA nanomaterials

“…2 F) [ 42 ], as well as functionalizing silica nanoparticles and carbon nanotubes with DNA (Fig. 2 G) [ 43 ]. These approaches allow for the construction of multifunctional DNA nanomaterials, expanding the scope of their applications in the biomedical field.…”

“…F Functionalization of CNT@DNA with DBP molecules containing specific FG and Attachment of multiple NPs onto a CNT [42]. G DNA-modified silica and carbon nanotubes were used to amplify the encoding plasmid of interlaced protein in a rolling circle [43] Hu and his team reported SiNP/CNT-DNA nanocomposites with high biocompatibility (Fig. 2C) [38].…”

Understanding the Structural Elasticity of RNA and DNA: All‐Atom Molecular Dynamics

“…Nucleic acids have essential roles in biological processes, including the replication and transcription of DNA, the reverse transcription of RNA viruses, and the gene regulation and expression of proteins. [1][2][3][4][5][6][7][8] For example, nucleic acids could kink with special protein residues and ions to perform various mechanical properties and regulate their elasticity of themselves in protein recognition. Thus, the structural elasticities are important for the nucleic acids, which are affected by many factors, such as the ionic DOI: 10.1002/adts.202200534 environments, [9][10][11][12][13] temperature, [14][15][16] and sequence orders.…”