M-TUBE enables large-volume bacterial gene delivery using a high-throughput microfluidic electroporation platform
<jats:p>Conventional cuvette-based and microfluidics-based electroporation approaches for bacterial gene delivery have distinct advantages, but they are typically limited to relatively small sample volumes, reducing their utility for applications requiring high throughput such as the generatio...
| Main Authors: | , , , , , |
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| Other Authors: | |
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2023
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| Online Access: | https://hdl.handle.net/1721.1/150803 |
| _version_ | 1826217810772099072 |
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| author | Huang, Po-Hsun Chen, Sijie Shiver, Anthony L Culver, Rebecca Neal Huang, Kerwyn Casey Buie, Cullen R |
| author2 | Massachusetts Institute of Technology. Department of Mechanical Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Mechanical Engineering Huang, Po-Hsun Chen, Sijie Shiver, Anthony L Culver, Rebecca Neal Huang, Kerwyn Casey Buie, Cullen R |
| author_sort | Huang, Po-Hsun |
| collection | MIT |
| description | <jats:p>Conventional cuvette-based and microfluidics-based electroporation approaches for bacterial gene delivery have distinct advantages, but they are typically limited to relatively small sample volumes, reducing their utility for applications requiring high throughput such as the generation of mutant libraries. Here, we present a scalable, large-scale bacterial gene delivery approach enabled by a disposable, user-friendly microfluidic electroporation device requiring minimal device fabrication and straightforward operation. We demonstrate that the proposed device can outperform conventional cuvettes in a range of situations, including across <jats:italic>Escherichia coli</jats:italic> strains with a range of electroporation efficiencies, and we use its large-volume bacterial electroporation capability to generate a library of transposon mutants in the anaerobic gut commensal <jats:italic>Bifidobacterium longum</jats:italic>.</jats:p> |
| first_indexed | 2024-09-23T17:09:31Z |
| format | Article |
| id | mit-1721.1/150803 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T17:09:31Z |
| publishDate | 2023 |
| publisher | Public Library of Science (PLoS) |
| record_format | dspace |
| spelling | mit-1721.1/1508032023-05-24T03:00:46Z M-TUBE enables large-volume bacterial gene delivery using a high-throughput microfluidic electroporation platform Huang, Po-Hsun Chen, Sijie Shiver, Anthony L Culver, Rebecca Neal Huang, Kerwyn Casey Buie, Cullen R Massachusetts Institute of Technology. Department of Mechanical Engineering <jats:p>Conventional cuvette-based and microfluidics-based electroporation approaches for bacterial gene delivery have distinct advantages, but they are typically limited to relatively small sample volumes, reducing their utility for applications requiring high throughput such as the generation of mutant libraries. Here, we present a scalable, large-scale bacterial gene delivery approach enabled by a disposable, user-friendly microfluidic electroporation device requiring minimal device fabrication and straightforward operation. We demonstrate that the proposed device can outperform conventional cuvettes in a range of situations, including across <jats:italic>Escherichia coli</jats:italic> strains with a range of electroporation efficiencies, and we use its large-volume bacterial electroporation capability to generate a library of transposon mutants in the anaerobic gut commensal <jats:italic>Bifidobacterium longum</jats:italic>.</jats:p> 2023-05-23T15:48:48Z 2023-05-23T15:48:48Z 2022 2023-05-23T15:41:12Z Article http://purl.org/eprint/type/JournalArticle https://hdl.handle.net/1721.1/150803 Huang, Po-Hsun, Chen, Sijie, Shiver, Anthony L, Culver, Rebecca Neal, Huang, Kerwyn Casey et al. 2022. "M-TUBE enables large-volume bacterial gene delivery using a high-throughput microfluidic electroporation platform." PLoS Biology, 20 (9). en 10.1371/JOURNAL.PBIO.3001727 PLoS Biology Creative Commons Attribution http://creativecommons.org/licenses/by/4.0/ application/pdf Public Library of Science (PLoS) PLOS |
| spellingShingle | Huang, Po-Hsun Chen, Sijie Shiver, Anthony L Culver, Rebecca Neal Huang, Kerwyn Casey Buie, Cullen R M-TUBE enables large-volume bacterial gene delivery using a high-throughput microfluidic electroporation platform |
| title | M-TUBE enables large-volume bacterial gene delivery using a high-throughput microfluidic electroporation platform |
| title_full | M-TUBE enables large-volume bacterial gene delivery using a high-throughput microfluidic electroporation platform |
| title_fullStr | M-TUBE enables large-volume bacterial gene delivery using a high-throughput microfluidic electroporation platform |
| title_full_unstemmed | M-TUBE enables large-volume bacterial gene delivery using a high-throughput microfluidic electroporation platform |
| title_short | M-TUBE enables large-volume bacterial gene delivery using a high-throughput microfluidic electroporation platform |
| title_sort | m tube enables large volume bacterial gene delivery using a high throughput microfluidic electroporation platform |
| url | https://hdl.handle.net/1721.1/150803 |
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