Toolbox for Exploring Modular Gene Regulation in Synthetic Biology Training.
Toolbox for Exploring Modular Gene Regulation in Synthetic Biology Training.
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用于探索合成生物学培训中的模块化基因调控的工具箱。
DOI：
10.1021/acssynbio.6b00057
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发表时间：
2016
影响因子：
4.7
通讯作者：
Chow,BrianY
中科院分区：
文献类型：
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作者：
Magaraci,MichaelS;Bermudez,JessicaG;Yogish,Deeksha;Pak,DanielH;Mollov,Viktor;Tycko,Josh;Issadore,David;Mannickarottu,SevileG;Chow,BrianY
我们报告了一个工具箱，用于探索遗传电路的模块化调整，该工具箱通过细菌菌株工程和可分发硬件开发的组合，专门针对STEM环境中的广泛部署进行了优化。16个基因开关的传递功能，编程表达GFP报告的（酰基-高丝氨酸内酯）AHL敏感的luxR转录激活剂的调节下，可以通过调节高/低程度的转录，翻译，和翻译后加工参数调谐。优化菌株以促进日常大规模制备和在室温下的可靠性能，以消除对温度控制装置的需要，温度控制装置既限制成本又限制空间。定制设计的，自动化的和网络启用的荧光记录系统允许AHL诱导的GFP表达在细菌平板上的延时成像，并具有实时远程数据访问，从而要求学员仅在实验设置时在场。当与符合经验数据的数学模型相结合时，该工具箱扩展了用于STEM培训的可靠合成生物学实验的可扩展性和范围。
We report a toolbox for exploring the modular tuning of genetic circuits, which has been specifically optimized for widespread deployment in STEM environments through a combination of bacterial strain engineering and distributable hardware development. The transfer functions of 16 genetic switches, programmed to express a GFP reporter under the regulation of the (acyl-homoserine lactone) AHL-sensitive luxR transcriptional activator, can be parametrically tuned by adjusting high/low degrees of transcriptional, translational, and post-translational processing. Strains were optimized to facilitate daily large-scale preparation and reliable performance at room temperature in order to eliminate the need for temperature controlled apparatuses, which are both cost-limiting and space-constraining. The custom-designed, automated, and web-enabled fluorescence documentation system allows time-lapse imaging of AHL-induced GFP expression on bacterial plates with real-time remote data access, thereby requiring trainees to only be present for experimental setup. When coupled with mathematical models in agreement with empirical data, this toolbox expands the scalability and scope of reliable synthetic biology experiments for STEM training.