Targeted Integration Through Transformation of Hydroxyurea-Arrested Cells.

Vasiliki Tsakraklides

Methods in molecular biology (Clifton, N.J.) 2021

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Homologous recombination is required to specifically target DNA to a desired genomic locus. Non-homologous end joining is the predominant form of recombination in Yarrowia lipolytica. Transformation of this organism with linear DNA therefore results in random integration of the introduced DNA into the genome. In this protocol, hydroxyurea-mediated cell cycle arrest is applied to significantly increase the rate of homologous recombination during transformation and enhance targeted integration.

Citation

Vasiliki Tsakraklides. Targeted Integration Through Transformation of Hydroxyurea-Arrested Cells. Methods in molecular biology (Clifton, N.J.). 2021;2307:139-145