Genetic Engineering of E. coli Nissle 1917 to Improve Transformation Efficiency
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2024-12
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Neilson, William
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Mansell, Thomas
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Shao, Zhengi
Wannemueler, Michael
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E. coli Nisslle 1917 is one of the most promising probiotic chassis for future work; however, it is an organism that struggles to be genetically modified without significant difficulty. In prior work, a directed evolution approach was taken to improve the rate at which electroporation could be performed in the cells. Here, we set out to determine if by copying the largest change in the genome observed in the evolved strain would result in a similar improvement in electroporation survival. To achieve this, a CRISPR Cas9 complex was used to target the carbamate kinases producing gene in the cell, deleting it in its entirety. While the adaptively evolved cells showed an upwards of ten-fold improvement in their transformation, there was no statistical difference in the transformation rate between the knockout strain and the control strain.
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creative component
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Attribution-NonCommercial 3.0 United States
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2024