Modular Synthetic Inverters from Zinc Finger Proteins and Small RNAs.
Modular Synthetic Inverters from Zinc Finger Proteins and Small RNAs.
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DOI：
10.1371/journal.pone.0149483
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Keasling JD
中科院分区：
文献类型：
--
作者：
Hsia J;Holtz WJ;Maharbiz MM;Arcak M;Keasling JD
合成的锌指蛋白（ZFP）可以靶向启动子DNA序列，抑制转录。小RNA（sRNA）与ZFP mRNA的结合产生了超灵敏的反应，以产生更高的有效希尔系数。在这里，我们结合了三个“现成的”ZFP和三个sRNA，在E.大肠杆菌中，并使用诱导折叠来量化它们的行为。我们发现ZFP和sRNA对诱导倍数的影响的一般顺序在组合这些部分时大多成立。然后，我们试图构建一个环形振荡器使用我们的新逆变器。我们所选择的部分表现不足以产生振荡，但我们包括未来的方向，以改进我们的工作在这里提出。
Synthetic zinc finger proteins (ZFPs) can be created to target promoter DNA sequences, repressing transcription. The binding of small RNA (sRNA) to ZFP mRNA creates an ultrasensitive response to generate higher effective Hill coefficients. Here we combined three “off the shelf” ZFPs and three sRNAs to create new modular inverters in E. coli and quantify their behavior using induction fold. We found a general ordering of the effects of the ZFPs and sRNAs on induction fold that mostly held true when combining these parts. We then attempted to construct a ring oscillator using our new inverters. Our chosen parts performed insufficiently to create oscillations, but we include future directions for improvement upon our work presented here.