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| Autori principali: | , , , , , , , |
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| Natura: | Preprint |
| Pubblicazione: |
2025
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| Soggetti: | |
| Accesso online: | https://arxiv.org/abs/2510.02468 |
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| _version_ | 1866915531095801856 |
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| author | Akh, Leyla Fajrial, Apresio K. Seelbinder, Benjamin Xu, Xin Tan, Wei Slansky, Jill Neu, Corey P. Ding, Xiaoyun |
| author_facet | Akh, Leyla Fajrial, Apresio K. Seelbinder, Benjamin Xu, Xin Tan, Wei Slansky, Jill Neu, Corey P. Ding, Xiaoyun |
| contents | 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. |
| format | Preprint |
| id |
arxiv_https___arxiv_org_abs_2510_02468 |
| institution | arXiv |
| publishDate | 2025 |
| record_format | arxiv |
| spellingShingle | Vector-free DNA transfection by nuclear envelope mechanoporation Akh, Leyla Fajrial, Apresio K. Seelbinder, Benjamin Xu, Xin Tan, Wei Slansky, Jill Neu, Corey P. Ding, Xiaoyun Biological Physics 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. |
| title | Vector-free DNA transfection by nuclear envelope mechanoporation |
| topic | Biological Physics |
| url | https://arxiv.org/abs/2510.02468 |