A Tunable and Expandable Transactivation System in Probiotic Yeast Saccharomyces boulardii.
A Tunable and Expandable Transactivation System in Probiotic Yeast Saccharomyces boulardii.
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DOI：
10.1021/acssynbio.1c00384
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发表时间：
2022-01-21
影响因子：
4.7
通讯作者：
Dantas, Gautam
中科院分区：
文献类型：
--
作者：
Kwak, Suryang;Mahmud, Bejan;Dantas, Gautam
精确的转录调节是开发具有可预测的可调功能的合成益生菌的关键要求。在这项研究中，一个可扩展的和可调的反式激活系统的构建和验证益生菌酵母布拉酵母。使用核酸酶无效的Cas9和支架RNA（scRNA）指导的调节使得能够在S.布拉地。由scRNA靶序列和核心GAL7启动子区域组成的合成启动子限制了来自天然半乳糖调节子的干扰。通过将新的靶序列引入启动子和scRNA，该系统很容易扩展。启动子和scRNA之间的互补性以及scRNA和转录激活因子之间的结合特异性充当了反式激活的两层正交性。此外，在诱导型启动子控制下的激活子表达使得能够通过化学诱导剂控制反式激活。所述系统具有使益生菌酵母工程化以更精确地执行治疗功能的潜力。
Precise transcriptional modulation is a key requirement for developing synthetic probiotics with predictably tunable functionalities. In this study, an expandable and tunable transactivation system was constructed and validated in probiotic yeast Saccharomyces boulardii. The use of nuclease-null Cas9 and scaffold RNA (scRNA) directed regulation enabled transactivation under the control of a synthetic promoter in S. boulardii. A synthetic promoter consisting of the scRNA target sequence and the core GAL7 promoter region restricted interference from the native galactose regulon. The system was readily expanded by introducing new target sequences to the promoter and scRNA. Complementarity between the promoter and scRNA, and binding specificity between scRNA and transcriptional activator, served as two layers of orthogonality of the transactivation. In addition, activator expression under the control of an inducible promoter enabled control of the transactivation via chemical inducer. The described system has potential to enable engineering of probiotic yeast to more precisely perform therapeutic functions.
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影响因子：
14.8
作者：
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通讯作者：
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影响因子：
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作者：
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1.2
作者：
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