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Adapting a CRISPR/Cas9 System to Study Electron Uptake in Shewanella oneidensis: Experimental Analysis and Proposals for Future Study

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Abstract

Though renewable energy production technologies have been improving over the last decade, energy storage technologies are still not up to par, hindering the widespread shift away from harmful fossil-based fuels. Biological processes like electron uptake can be repurposed for energy storage through rewired carbon fixation. One organism of interest for this task is Shewanella oneidensis, but the molecular mechanisms of its electron uptake pathway are not fully characterized or understood, and it lacks a robust genetic toolkit to enable the full illumination of its potential. This paper explores a system capable of generating single-gene knockouts quickly and precisely in S. oneidensis by using single-stranded DNA oligonucleotide recombineering coupled with CRISPR/Cas9 counterselection. This paper also explores future possibilities involving the scaling up of this system to multiple genes and the addition of this system to a genetic toolkit for S. oneidensis.

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2020-06-01

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bioenergy; bioengineering; Shewanella; electron uptake; CRISPR

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Government Document

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Attribution-NonCommercial-NoDerivatives 4.0 International

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dissertation or thesis

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