Formulation of DNA Nanocomposites: Towards Functional Materials for Protein Expression.
Formulation of DNA Nanocomposites: Towards Functional Materials for Protein Expression.
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DNA纳米复合材料的配方：朝向蛋白质表达的功能材料。
DOI：
10.3390/polym13152395
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发表时间：
2021-07-21
期刊：
影响因子：
5
通讯作者：
Niemeyer CM
中科院分区：
文献类型：
--
作者：
Schipperges A;Hu Y;Moench S;Weigel S;Reith J;Ordoñez-Rueda D;Rabe KS;Niemeyer CM
DNA 水凝胶是一类新兴材料，在从组织工程到靶向药物输送和无细胞蛋白质合成 (CFPS) 等众多生物技术应用中具有广阔的前景。除了可用于指导生物系统的 DNA 分子可编程性之外，DNA 材料的配方（例如块状水凝胶或微凝胶）也与特定应用相关。为了推进这一研究领域的知识水平，目前的工作探讨了最近开发的一类复杂 DNA 纳米复合材料的范围，该复合材料是通过 DNA 功能化二氧化硅纳米颗粒 (SiNP) 和碳纳米管 (CNT) 的 RCA 聚合合成的。 SiNP/CNT-DNA 复合材料以散装材料和微凝胶的形式生产，其中含有带有荧光标记蛋白可转录遗传信息的质粒。使用共聚焦显微镜和流式细胞术，我们发现这些材料被各种真核细胞系非常有效地吸收，这些细胞系能够继续分裂，同时摄入的材料均匀地分布到子细胞中。然而，细胞内没有发生编码蛋白质的表达。尽管即使在使用真核细胞裂解物的 CFPS 程序中，微凝胶也不会诱导标记蛋白的产生，但大量复合材料被证明是 CFPS 的有效模板。这项工作有助于理解 DNA 复合材料和功能性细胞机制之间的分子相互作用。讨论了使用此类材料对 CFPS 程序的影响。
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.
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