Improved Gene Targeting through Cell Cycle Synchronization
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Tsakraklides-2015-Improved Gene Target.pdf
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411.54 KB
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Author(s)
Tsakraklides, Vasiliki
Brevnova, Elena
Stephanopoulos, Gregory
Shaw, A. Joe
Date Issued
July 2015
Journal
PLOS ONE
Publisher
Public Library of Science
Citation
Tsakraklides, Vasiliki, Elena Brevnova, Gregory Stephanopoulos, and A. Joe Shaw. “Improved Gene Targeting through Cell Cycle Synchronization.” Edited by Alvaro Galli. PLoS ONE 10, no. 7 (July 20, 2015): e0133434.
Version
Final published version
Abstract
Gene targeting is a challenge in organisms where non-homologous end-joining is the predominant form of recombination. We show that cell division cycle synchronization can be applied to significantly increase the rate of homologous recombination during transformation. Using hydroxyurea-mediated cell cycle arrest, we obtained improved gene targeting rates in Yarrowia lipolytica, Arxula adeninivorans, Saccharomyces cerevisiae, Kluyveromyces lactis and Pichia pastoris demonstrating the broad applicability of the method. Hydroxyurea treatment enriches for S-phase cells that are active in homologous recombination and enables previously unattainable genomic modifications.
MIT Department
Massachusetts Institute of Technology. Department of Chemical Engineering
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Creative Commons Attribution
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DOI of Published Version
https://doi.org/10.1371/journal.pone.0133434
