Improved Gene Targeting through Cell Cycle Synchronization.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 26192309.
- Also identified by DOI 10.1371/journal.pone.0133434 and PMC identifier 4507847.
- Licence recorded as CC BY.
- The licence permits redistribution, so the abstract is shown in full and the full text is available from the publisher.
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.
Medical subject headings
- Cell Cycle
- Gene Targeting