Department of Chemical Engineering, University of California Santa Barbara, Santa Barbara, CA, 93106, USA.
Nat Commun. 2020 Oct 29;11(1):5459. doi: 10.1038/s41467-020-19271-1.
Biocontainment systems are needed to neutralize genetically modified organisms (GMOs) that pose ecological threats outside of controlled environments. In contrast, benign selection markers complement GMOs with reduced fitness. Benign selection agents serve as alternatives to antibiotics, which are costly and risk spread of antibiotic resistance. Here, we present a yeast biocontainment strategy leveraging engineered fluoride sensitivity and DNA vectors enabling use of fluoride as a selection agent. The biocontainment system addresses the scarcity of platforms available for yeast despite their prevalent use in industry and academia. In the absence of fluoride, the biocontainment strain exhibits phenotypes nearly identical to those of the wildtype strain. Low fluoride concentrations severely inhibit biocontainment strain growth, which is restored upon introduction of fluoride-based vectors. The biocontainment strategy is stringent, easily implemented, and applicable to several eukaryotes. Further, the DNA vectors enable genetic engineering at reduced costs and eliminate risks of propagating antibiotic resistance.
生物 containment 系统需要用来中和在受控环境之外具有生态威胁的转基因生物（GMO）。相比之下，良性选择标记物使具有降低适应性的 GMO 得到补充。良性选择剂可替代抗生素，因为抗生素成本高且有抗生素耐药性传播的风险。在这里，我们提出了一种酵母生物 containment 策略，利用工程化的氟化物敏感性和 DNA 载体，使氟化物可用作选择剂。该生物 containment 系统解决了尽管酵母在工业和学术界中广泛使用，但可用平台稀缺的问题。在没有氟化物的情况下，生物 containment 菌株表现出的表型与野生型菌株几乎相同。低浓度的氟化物严重抑制生物 containment 菌株的生长，而在引入基于氟化物的载体后，生长得到恢复。该生物 containment 策略严格、易于实施，并且适用于几种真核生物。此外，DNA 载体可降低成本进行基因工程，并消除传播抗生素耐药性的风险。
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