Intrinsic biocontainment: Multiplex genome safeguards combine transcriptional and recombinational control of essential yeast genes

Yizhi Cai, Neta Agmon, Woo Jin Choi, Alba Ubide, Giovanni Stracquadanio, Katrina Caravelli, Haiping Hao, Joel S. Bader, Jef D. Boeke

    Research output: Contribution to journalArticle

    Abstract

    Biocontainment may be required in a wide variety of situations such as work with pathogens, field release applications of engineered organisms, and protection of intellectual properties. Here, we describe the control of growth of the brewer's yeast, Saccharomyces cerevisiae, using both transcriptional and recombinational "safeguard" control of essential gene function. Practical biocontainment strategies dependent on the presence of small molecules require them to be active at very low concentrations, rendering them inexpensive and difficult to detect. Histone genes were controlled by an inducible promoter and controlled by 30 nM estradiol. The stability of the engineered genes was separately regulated by the expression of a site-specific recombinase. The combined frequency of generating viable derivativeswhen both systemswere active was below detection (<10-10), consistent with their orthogonal nature and the individual escape frequencies of <10-6. Evaluation of escaper mutants suggests strategies for reducing their emergence. Transcript profiling and growth test suggest high fitness of safeguarded strains, an important characteristic for wide acceptance.

    Original languageEnglish (US)
    Pages (from-to)1803-1808
    Number of pages6
    JournalProceedings of the National Academy of Sciences of the United States of America
    Volume112
    Issue number6
    DOIs
    StatePublished - Feb 10 2015

    Keywords

    • Biosafety
    • Biotechnology
    • Saccharomyces cerevisiae

    ASJC Scopus subject areas

    • General

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