Transient Silencing of DNA Repair Genes Improves Targeted Gene Integration in the Filamentous Fungus Trichoderma reesei

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    Abstract

    Trichoderma reesei is a filamentous fungus that is used worldwide to produce industrial enzymes. Industrial strains have traditionally been created though systematic strain improvement using mutagenesis and screening approaches. It is also desirable to specifically manipulate the genes of the organism to further improve and to modify the strain. Targeted integration in filamentous fungi is typically hampered by very low frequencies of homologous recombination. To address this limitation, we have developed a simple transient method for silencing genes in T. reesei. Using gene-specific small interfering RNAs (siRNAs) targeted to mus53, we could achieve up to 90% knockdown of mus53 mRNA. As a practical example, we demonstrated that transient silencing of DNA repair genes significantly improved homologous integration of DNA at a specific locus in a standard protoplast transformation. The best transient silencing of mus53 with siRNAs in protoplasts could achieve up to 59% marker gene integration.

    Original languageEnglish
    Article numbere00535-17
    Number of pages11
    JournalApplied and Environmental Microbiology
    Volume83
    Issue number15
    DOIs
    Publication statusPublished - 2017
    MoE publication typeA1 Journal article-refereed

    Keywords

    • Trichoderma reesei
    • filamentous fungi
    • gene silencing
    • industrial biotechnology
    • siRNA

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