| Autor: |
Kosuke Iwai, Maren Wehrs, Megan Garber, Jess Sustarich, Lauren Washburn, Zachary Costello, Peter W. Kim, David Ando, William R. Gaillard, Nathan J. Hillson, Paul D. Adams, Aindrila Mukhopadhyay, Hector Garcia Martin, Anup K. Singh |
| Jazyk: |
angličtina |
| Rok vydání: |
2022 |
| Předmět: |
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| Zdroj: |
Microsystems & Nanoengineering, Vol 8, Iss 1, Pp 1-10 (2022) |
| Druh dokumentu: |
article |
| ISSN: |
2055-7434 |
| DOI: |
10.1038/s41378-022-00357-3 |
| Popis: |
Abstract We present a droplet-based microfluidic system that enables CRISPR-based gene editing and high-throughput screening on a chip. The microfluidic device contains a 10 × 10 element array, and each element contains sets of electrodes for two electric field-actuated operations: electrowetting for merging droplets to mix reagents and electroporation for transformation. This device can perform up to 100 genetic modification reactions in parallel, providing a scalable platform for generating the large number of engineered strains required for the combinatorial optimization of genetic pathways and predictable bioengineering. We demonstrate the system’s capabilities through the CRISPR-based engineering of two test cases: (1) disruption of the function of the enzyme galactokinase (galK) in E. coli and (2) targeted engineering of the glutamine synthetase gene (glnA) and the blue-pigment synthetase gene (bpsA) to improve indigoidine production in E. coli. |
| Databáze: |
Directory of Open Access Journals |
| Externí odkaz: |
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