Journal
NUCLEIC ACIDS RESEARCH
Volume 39, Issue 3, Pages -Publisher
OXFORD UNIV PRESS
DOI: 10.1093/nar/gkq1070
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Funding
- JFE 21st Century Foundation
- Ministry of Education, Culture, Sports, Science and Technology, Japan
- Futaba Electronics Memorial Foundation
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The development of genetic switches and their integrated forms (genetic circuits) with desired specifications/functions is key for success in synthetic biology. Due to the difficulty in rational design, genetic switches and circuits with desirable specifications are mostly obtained by directed evolution. Based on a virus-derived nucleotide kinase as a single-gene dual selector, we constructed a robust, efficient and stringent selection system for genetic switches. This method exhibited unprecedented enrichment efficacy (> 30 000-fold) of functional switches from non-functional ones in a single selection cycle. In addition, negative (OFF) selection was exceptionally stringent, allowing the rapid and efficient selection of non-leaky from leaky circuits.
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