Multiplex Genome Editing by Natural Transformation (MuGENT) for Synthetic Biology in <i>Vibrio natriegens</i>

Published on 2018-02-10T00:13:41Z (GMT) by
<i>Vibrio natriegens</i> has recently emerged as an alternative to <i>Escherichia coli</i> for molecular biology and biotechnology, but low-efficiency genetic tools hamper its development. Here, we uncover how to induce natural competence in <i>V. natriegens</i> and describe methods for multiplex genome editing by natural transformation (MuGENT). MuGENT promotes integration of multiple genome edits at high-efficiency on unprecedented time scales. Also, this method allows for generating highly complex mutant populations, which can be exploited for metabolic engineering efforts. As a proof-of-concept, we attempted to enhance production of the value added chemical poly-β-hydroxybutyrate (PHB) in <i>V. natriegens</i> by targeting the expression of nine genes involved in PHB biosynthesis <i>via</i> MuGENT. Within 1 week, we isolated edited strains that produced ∼100 times more PHB than the parent isolate and ∼3.3 times more than a rationally designed strain. Thus, the methods described here should extend the utility of this species for diverse academic and industrial applications.

Cite this collection

Dalia, Triana N.; Hayes, Chelsea A.; Stolyar, Sergey; J. Marx, Christopher; McKinlay, James B.; B. Dalia, Ankur (2018): Multiplex Genome Editing by Natural Transformation

(MuGENT) for Synthetic Biology in Vibrio natriegens. ACS Publications. Collection.