Department of Biochemistry and Molecular Biology, Iowa State University, Ames, IA, 50011, USA.
Department of Oncology, University of Wisconsin-Madison, Madison, WI, 53705, USA.
Sci Rep. 2024 Nov 9;14(1):27364. doi: 10.1038/s41598-024-78399-y.
Self-assembling DNA nanoparticles have the potential to significantly advance the targeted delivery of molecular cargo owing to their chemical and architectural flexibility. Recently, it has been demonstrated that the genetic code embedded in DNA nanoparticles produced by the method of DNA origami or related techniques can be recognized and copied by RNA polymerase in vitro. Further, sculpted DNA nanoparticles can serve as a substrate for Cas9-mediated gene modification and gene expression in cell culture. In the present study, we further investigate the ability of DNA origami nanoparticles to be expressed in a human cell line with emphasis on the impact of single-stranded DNA (ssDNA) domains and the contributions of the architectural disposition of genetic control elements, namely promoter and enhancer sequences. Our findings suggest that while cells possess the remarkable capability to express genes within highly folded architectures, the presence and relative density and location of ssDNA domains appears to influence overall levels of gene expression. These results suggest that it may be possible to nuance folded DNA nanoparticle architecture to regulate the rate and/or level of gene expression. Considering the highly malleable architecture and chemistry of self-assembling DNA nanoparticles, these findings motivate further exploration of their potential as an economic nanotechnology platform for targeted gene editing, nucleic acid-based vaccines, and related biotherapeutic applications.
自组装 DNA 纳米颗粒由于其化学和结构的灵活性，有可能显著推进分子货物的靶向递送。最近已经证明，通过 DNA 折纸术或相关技术产生的 DNA 纳米颗粒中嵌入的遗传密码可以被体外的 RNA 聚合酶识别和复制。此外，雕刻 DNA 纳米颗粒可以作为 Cas9 介导的基因修饰和细胞培养中基因表达的底物。在本研究中，我们进一步研究了 DNA 折纸纳米颗粒在人细胞系中表达的能力，重点研究了单链 DNA（ssDNA）结构域的影响以及遗传调控元件（如启动子和增强子序列）的结构排列的贡献。我们的研究结果表明，尽管细胞具有在高度折叠的结构中表达基因的非凡能力，但 ssDNA 结构域的存在、相对密度和位置似乎会影响基因表达的整体水平。这些结果表明，可能可以通过调整折叠 DNA 纳米颗粒的结构来调节基因表达的速度和/或水平。考虑到自组装 DNA 纳米颗粒高度可塑的结构和化学性质，这些发现促使进一步探索它们作为一种经济的纳米技术平台，用于靶向基因编辑、基于核酸的疫苗和相关的生物治疗应用的潜力。
