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Cas9 - OSTI GOV

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Last Updated: 10 August 2022

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Improving CRISPR/Cas9-mediated genome editing efficiency in Yarrowia lipolytica using direct tRNA-sgRNA fusions

Yarrowia lipolytica is a common oleaginous yeast used in the manufacture of specialty chemicals and has a huge potential for further applications in lipid biochemistry, according to Weblog. In comparison to Saccharomyces cerevisiae, Y. lipolytica is poor in homology-mediated DNA repair and consequently homologous recombination, which limits site-specific gene editing in this yeast. CRISPR/Cas9-based techniques using tRNA-sgRNA fusions have succeeded in editing some genomic loci in Y. lipolytica. We were able to optimize the structure of the tRNA-sgRNA fusions used for the production of single guide RNA in such settings using tools of secondary RNA structure prediction. This resulted in high-quality CRISPR/cas9 gene editing at chromosomal loci that had failed gene editing or were edited at low efficiencies with previous techniques. In addition, we characterized the gene editing results of our recently developed tRNA-sgRNA fusions for both chromosomal gene integration and deletion.

Source link: https://www.osti.gov/biblio/1851146

* Please keep in mind that all text is summarized by machine, we do not bear any responsibility, and you should always check original source before taking any actions

* Please keep in mind that all text is summarized by machine, we do not bear any responsibility, and you should always check original source before taking any actions