通过核膜机械穿孔实现无载体DNA转染
Vector-free DNA transfection by nuclear envelope mechanoporation
AI总结：
针对现有核酸递送方法的载体缺陷及机械法穿孔的局限，本研究提出集成高纵横比纳米刺针的微流控装置，可直接向细胞核递送裸DNA，实现高存活率、高表达速度且通量显著高于显微注射，有望用于快速制备大量基因工程细胞治疗疾病。
摘要：
细胞基因工程在治疗不治之症方面有诸多应用。质粒DNA因成本低、稳定性高，是细胞工程中常用的核酸选择。然而，质粒DNA必须避开细胞质中的保护机制，进入细胞核才能进行翻译。现有的许多核酸递送方法，如化学递送法和病毒递送法，都存在由核酸载体本身引发的缺陷。机械方法为核酸递送提供了另一种思路，它通过物理方式在细胞上制造开口以递送货物。但在大多数系统中，细胞膜开口过小，无法递送大分子货物，或者穿孔过程会导致细胞存活率低。本研究提出一种集成了高纵横比纳米结构的微流控装置，可反复刺破细胞膜和核膜。这些尖端锋利的纳米刺针（nanolancets）能足够深地穿透细胞，将货物直接递送至细胞核，同时仍能让细胞在处理后恢复。我们证明了该装置能够向多种细胞类型递送货物，同时保持高存活率。随后，我们证实了裸DNA直接递送至细胞核可使质粒DNA快速启动表达，其表达速度与显微注射相当，但通量显著更高。我们设想该装置可作为一种工具，快速制备大量基因工程细胞以治疗多种疾病。
英文摘要：
Genetic engineering of cells has a range of applications in treating incurable diseases. Plasmid DNA is a popular choice of nucleic acid for cell engineering due to its low cost and stability. However, plasmid DNA must survive the protective mechanisms present in the cell's cytoplasm to enter the nucleus for translation. Many of the existing methods for nucleic acid delivery, such as chemical-based and virus-based delivery, suffer from drawbacks induced by the nucleic acid carrier itself. Mechanical methods present an alternative to nucleic acid carriers by physically producing openings in the cell to deliver cargos. However, in most systems, the cell membrane openings are too small to deliver large cargos, or the poration process leads to low cell viability. In this study, we present a microfluidic device with integrated high aspect ratio nanostructures that repeatably rupture the cell membrane and nuclear envelope. These sharp-tipped nanolancets penetrate the cell deep enough to allow direct delivery of cargos into the nucleus, but still allow for cell recovery after treatment. We show the device's ability to deliver cargo to a variety of cell types while maintaining high viability. Then, we demonstrate the rapid onset of plasmid DNA expression that results from direct nuclear delivery of naked DNA, showing expression speeds comparable to microinjection, but with significantly greater throughput. We envision the use of this device as a tool to quickly produce high quantities of genetically engineered cells to treat a myriad of diseases.
机构、总结、中文摘要均由AI分析而来，可能存在错误，请以论文正文为准。
