Improved Gene Targeting through Cell Cycle Synchronization.
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...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2015-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC4507847?pdf=render |
| _version_ | 1828744292162600960 |
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| author | Vasiliki Tsakraklides Elena Brevnova Gregory Stephanopoulos A Joe Shaw |
| author_facet | Vasiliki Tsakraklides Elena Brevnova Gregory Stephanopoulos A Joe Shaw |
| author_sort | Vasiliki Tsakraklides |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-13T01:54:25Z |
| format | Article |
| id | doaj.art-d0adeac608384b889a3a68b67f65af1f |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-04-13T01:54:25Z |
| publishDate | 2015-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-d0adeac608384b889a3a68b67f65af1f2022-12-22T03:07:48ZengPublic Library of Science (PLoS)PLoS ONE1932-62032015-01-01107e013343410.1371/journal.pone.0133434Improved Gene Targeting through Cell Cycle Synchronization.Vasiliki TsakraklidesElena BrevnovaGregory StephanopoulosA Joe ShawGene 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.http://europepmc.org/articles/PMC4507847?pdf=render |
| spellingShingle | Vasiliki Tsakraklides Elena Brevnova Gregory Stephanopoulos A Joe Shaw Improved Gene Targeting through Cell Cycle Synchronization. PLoS ONE |
| title | Improved Gene Targeting through Cell Cycle Synchronization. |
| title_full | Improved Gene Targeting through Cell Cycle Synchronization. |
| title_fullStr | Improved Gene Targeting through Cell Cycle Synchronization. |
| title_full_unstemmed | Improved Gene Targeting through Cell Cycle Synchronization. |
| title_short | Improved Gene Targeting through Cell Cycle Synchronization. |
| title_sort | improved gene targeting through cell cycle synchronization |
| url | http://europepmc.org/articles/PMC4507847?pdf=render |
| work_keys_str_mv | AT vasilikitsakraklides improvedgenetargetingthroughcellcyclesynchronization AT elenabrevnova improvedgenetargetingthroughcellcyclesynchronization AT gregorystephanopoulos improvedgenetargetingthroughcellcyclesynchronization AT ajoeshaw improvedgenetargetingthroughcellcyclesynchronization |