Rational Design of Evolutionarily Stable Microbial Kill Switches
| Autor: | Pamela A. Silver, Elizabeth Redfield, Jeffrey C. Way, Samuel O’Keefe, Finn Stirling, Lisa Bitzan, John Oliver |
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| Rok vydání: | 2017 |
| Předmět: |
DNA
Bacterial 0301 basic medicine Mice Inbred BALB C education.field_of_study Escherichia coli K12 Escherichia coli Proteins 030106 microbiology Population Evolutionary stability Rational design Cell Biology Computational biology Biology Microbiology Mice 03 medical and health sciences Synthetic biology 030104 developmental biology Animals Directed Molecular Evolution Antitoxin Kill switch education Molecular Biology |
| Zdroj: | Molecular Cell. 68:686-697.e3 |
| ISSN: | 1097-2765 |
| DOI: | 10.1016/j.molcel.2017.10.033 |
| Popis: | Summary The evolutionary stability of synthetic genetic circuits is key to both the understanding and application of genetic control elements. One useful but challenging situation is a switch between life and death depending on environment. Here are presented "essentializer" and "cryodeath" circuits, which act as kill switches in Escherichia coli . The essentializer element induces cell death upon the loss of a bi-stable cI/Cro memory switch. Cryodeath makes use of a cold-inducible promoter to express a toxin. We employ rational design and a toxin/antitoxin titering approach to produce and screen a small library of potential constructs, in order to select for constructs that are evolutionarily stable. Both kill switches were shown to maintain functionality in vitro for at least 140 generations. Additionally, cryodeath was shown to control the growth environment of a population, with an escape frequency of less than 1 in 10 5 after 10 days of growth in the mammalian gut. |
| Databáze: | OpenAIRE |
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