Deletion of DNA ligase IV homolog confers higher gene targeting efficiency on homologous recombination in Komagataella phaffii

FEMS Yeast Research
Yoichiro ItoAkihiko Kondo

Abstract

The non-conventional yeast Komagataella phaffii, formerly Pichia pastoris, is a popular host for recombinant protein production. The relatively lower gene targeting efficiency observed in this species occurs due to high levels of non-homologous recombination activity. In the current study, we explored the function of the K. phaffii homolog of DNA ligase IV (Dnl4p) by creating a DNL4-disrupted strain. To assess the roles of non-homologous end joining (NHEJ)-related proteins in this species, strains deleted for either or both genes encoding Dnl4p or the telomeric Ku complex subunit (Ku70p) were generated. These deletions were constructed by either of two distinct marker-recycling methods (yielding either a seamless gene deletion or a Cre-loxP-mediated gene deletion). The resulting dnl4- and/or ku70-deleted K. phaffii strains were used to evaluate gene targeting efficiency in gene knock-out and gene knock-in experiments. The Dnl4p-defective strain showed improved gene targeting efficiency for homologous recombination compared to the wild-type and Ku70p-deffective strains. The dnl4 ku70 double knock-out strain exhibited a further improvement in gene targeting efficiency. Thus, the K. phaffii dnl4 and dnl4 ku70 deletion strains are ...Continue Reading

References

Oct 1, 1979·Proceedings of the National Academy of Sciences of the United States of America·S Scherer, R W Davis
Dec 1, 1985·Molecular and Cellular Biology·J M CreggK R Madden
Jul 31, 1997·Nature·T E WilsonM R Lieber
Apr 28, 2004·FEMS Microbiology Reviews·U Klinner, B Schäfer
Jul 30, 2004·Yeast·Rolf KooistraH Yde Steensma
Aug 10, 2004·Proceedings of the National Academy of Sciences of the United States of America·Yuuko NinomiyaHirokazu Inoue
Nov 16, 2005·Annual Review of Genetics·James M DaleyThomas E Wilson
Sep 28, 2006·Proceedings of the National Academy of Sciences of the United States of America·Kazuma IshibashiHirokazu Inoue
Jun 10, 2008·Nucleic Acids Research·Franz S HartnerAnton Glieder
Jul 31, 2008·Microbial Cell Factories·Hans MarxMichael Sauer
May 26, 2009·Nature Biotechnology·Kristof De SchutterNico Callewaert
Jan 7, 2011·FEMS Yeast Research·Rongqing PanXin Yan
Apr 19, 2011·Applied and Environmental Microbiology·Xiulin QinJu Chu
May 18, 2011·Journal of Biotechnology·Andreas KüberlHarald Pichler
Jun 15, 2011·Applied and Environmental Microbiology·Nadja Graf, Josef Altenbuchner
Jan 26, 2013·Microbial Cell Factories·Roland PrielhoferDiethard Mattanovich
Feb 5, 2013·Future Microbiology·Brigitte GasserDiethard Mattanovich
Nov 6, 2013·ACS Synthetic Biology·Thomas VoglAnton Glieder
Apr 20, 2014·Applied Microbiology and Biotechnology·Mudassar AhmadHelmut Schwab
Dec 3, 2015·Microbial Cell Factories·Brigitte GasserDiethard Mattanovich
Apr 17, 2016·Journal of Biotechnology·Lukas SturmbergerAnton Glieder
Aug 9, 2016·BMC Genomics·Kerry R LoveJ Christopher Love
May 29, 2018·Applied and Environmental Microbiology·Yasuyuki NakamuraAkihiko Kondo

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Citations

Aug 27, 2021·Frontiers in Bioengineering and Biotechnology·Gita NaseriChristoph Arenz

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