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

Cai, Y and Agmon, N and Choi, WJ and Ubide, A and Stracquadanio, G and Caravelli, K and Hao, H and Bader, JS and Boeke, JD (2015) 'Intrinsic biocontainment: Multiplex genome safeguards combine transcriptional and recombinational control of essential yeast genes.' Proceedings of the National Academy of Sciences of the United States of America, 112 (6). 1803 - 1808. ISSN 0027-8424

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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.

Item Type: Article
Subjects: Q Science > QA Mathematics > QA75 Electronic computers. Computer science
Q Science > QD Chemistry
Divisions: Faculty of Science and Health > Computer Science and Electronic Engineering, School of
Depositing User: Giovanni Stracquadanio
Date Deposited: 16 Jan 2017 12:30
Last Modified: 17 Aug 2017 17:20
URI: http://repository.essex.ac.uk/id/eprint/18692

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