High-throughput nuclear delivery and rapid expression of DNA via mechanical and electrical cell-membrane disruption
Nuclear transfection of DNA into mammalian cells is challenging yet critical for many biological and medical studies. Here, by combining cell squeezing and electric-field-driven transport in a device that integrates microfluidic channels with constrictions and microelectrodes, we demonstrate nuclear...
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| Format: | Article |
| Language: | en_US |
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2017
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| Online Access: | http://hdl.handle.net/1721.1/111022 https://orcid.org/0000-0003-4252-9335 https://orcid.org/0000-0003-4112-6622 https://orcid.org/0000-0002-9016-5962 https://orcid.org/0000-0003-4255-0492 https://orcid.org/0000-0001-7192-580X |
| _version_ | 1826199522568568832 |
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| author | Ding, Xiaoyun Sean Stewart, Martin P Sharei, Armon Reza Weaver, James C Langer, Robert S Jensen, Klavs F |
| author2 | Harvard University--MIT Division of Health Sciences and Technology |
| author_facet | Harvard University--MIT Division of Health Sciences and Technology Ding, Xiaoyun Sean Stewart, Martin P Sharei, Armon Reza Weaver, James C Langer, Robert S Jensen, Klavs F |
| author_sort | Ding, Xiaoyun Sean |
| collection | MIT |
| description | Nuclear transfection of DNA into mammalian cells is challenging yet critical for many biological and medical studies. Here, by combining cell squeezing and electric-field-driven transport in a device that integrates microfluidic channels with constrictions and microelectrodes, we demonstrate nuclear delivery of plasmid DNA within 1 h after treatment—the most rapid DNA expression in a high-throughput setting (up to millions of cells per minute per device). Passing cells at high speed through microfluidic constrictions smaller than the cell diameter mechanically disrupts the cell membrane, allowing a subsequent electric field to further disrupt the nuclear envelope and drive DNA molecules into the cytoplasm and nucleus. By tracking the localization of the endosomal sorting complex required for transport III protein CHMP4B (charged multivesicular body protein 4B), we show that the integrity of the nuclear envelope is recovered within 15 minutes of treatment. We also provide insight into subcellular delivery by comparing the performance of the disruption-and-field-enhanced method with those of conventional chemical, electroporation and manual-injection systems. |
| first_indexed | 2024-09-23T11:21:41Z |
| format | Article |
| id | mit-1721.1/111022 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T11:21:41Z |
| publishDate | 2017 |
| record_format | dspace |
| spelling | mit-1721.1/1110222022-10-01T03:06:00Z High-throughput nuclear delivery and rapid expression of DNA via mechanical and electrical cell-membrane disruption Ding, Xiaoyun Sean Stewart, Martin P Sharei, Armon Reza Weaver, James C Langer, Robert S Jensen, Klavs F Harvard University--MIT Division of Health Sciences and Technology Massachusetts Institute of Technology. Department of Chemical Engineering Koch Institute for Integrative Cancer Research at MIT Jensen, Klavs F. Ding, Xiaoyun Sean Stewart, Martin P Sharei, Armon Reza Weaver, James C Langer, Robert S Jensen, Klavs F Nuclear transfection of DNA into mammalian cells is challenging yet critical for many biological and medical studies. Here, by combining cell squeezing and electric-field-driven transport in a device that integrates microfluidic channels with constrictions and microelectrodes, we demonstrate nuclear delivery of plasmid DNA within 1 h after treatment—the most rapid DNA expression in a high-throughput setting (up to millions of cells per minute per device). Passing cells at high speed through microfluidic constrictions smaller than the cell diameter mechanically disrupts the cell membrane, allowing a subsequent electric field to further disrupt the nuclear envelope and drive DNA molecules into the cytoplasm and nucleus. By tracking the localization of the endosomal sorting complex required for transport III protein CHMP4B (charged multivesicular body protein 4B), we show that the integrity of the nuclear envelope is recovered within 15 minutes of treatment. We also provide insight into subcellular delivery by comparing the performance of the disruption-and-field-enhanced method with those of conventional chemical, electroporation and manual-injection systems. National Institutes of Health (U.S.) (R01GM101420-01A1) 2017-08-28T17:18:50Z 2017-08-28T17:18:50Z 2017-03 2016-05 Article http://purl.org/eprint/type/JournalArticle 2157-846X http://hdl.handle.net/1721.1/111022 Ding, Xiaoyun, et al. “High-Throughput Nuclear Delivery and Rapid Expression of DNA via Mechanical and Electrical Cell-Membrane Disruption.” Nature Biomedical Engineering 1, 3 (March 2017): 0039 © 2017 Macmillan Publishers Limited, part of Springer Nature https://orcid.org/0000-0003-4252-9335 https://orcid.org/0000-0003-4112-6622 https://orcid.org/0000-0002-9016-5962 https://orcid.org/0000-0003-4255-0492 https://orcid.org/0000-0001-7192-580X en_US http://dx.doi.org/10.1038/s41551-017-0039 Nature Biomedical Engineering Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf Prof. Langer via Erja Kajosalo |
| spellingShingle | Ding, Xiaoyun Sean Stewart, Martin P Sharei, Armon Reza Weaver, James C Langer, Robert S Jensen, Klavs F High-throughput nuclear delivery and rapid expression of DNA via mechanical and electrical cell-membrane disruption |
| title | High-throughput nuclear delivery and rapid expression of DNA via mechanical and electrical cell-membrane disruption |
| title_full | High-throughput nuclear delivery and rapid expression of DNA via mechanical and electrical cell-membrane disruption |
| title_fullStr | High-throughput nuclear delivery and rapid expression of DNA via mechanical and electrical cell-membrane disruption |
| title_full_unstemmed | High-throughput nuclear delivery and rapid expression of DNA via mechanical and electrical cell-membrane disruption |
| title_short | High-throughput nuclear delivery and rapid expression of DNA via mechanical and electrical cell-membrane disruption |
| title_sort | high throughput nuclear delivery and rapid expression of dna via mechanical and electrical cell membrane disruption |
| url | http://hdl.handle.net/1721.1/111022 https://orcid.org/0000-0003-4252-9335 https://orcid.org/0000-0003-4112-6622 https://orcid.org/0000-0002-9016-5962 https://orcid.org/0000-0003-4255-0492 https://orcid.org/0000-0001-7192-580X |
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