Construction of Two-Input Logic Gates Using Transcriptional Interference
Construction of Two-Input Logic Gates Using Transcriptional Interference
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DOI：
10.1021/acssynbio.9b00321
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发表时间：
2019-10-01
影响因子：
4.7
通讯作者：
Chatterjee, Anushree
中科院分区：
文献类型：
--
作者：
Bordoy, Antoni E.;O'Connor, Nolan J.;Chatterjee, Anushree
转录干扰（TI）已被证明通过不同的分子机制在DNA水平上调节基因表达。转录RNA聚合酶可能遇到的DNA上存在的障碍，例如DNA结合蛋白或另一种RNA聚合酶，可以导致TI，导致转录终止，从而降低基因表达。然而，TI作为一种新的策略，工程复杂的基因表达模块的潜力还没有得到充分的探讨。在这里，我们创建了一系列双输入遗传设备，这些设备使用路障蛋白的存在来控制基因表达，并使用实验和数学建模方法分析了它们的行为。我们探讨了多个特征如何影响遗传装置的反应，这些遗传装置被设计成像AND、OR或单输入逻辑门一样。我们发现，路障蛋白的解离常数，诱导剂激活的启动子和操作位点，和串联启动子之间的距离调整门的行为。这项工作突出了在微妙不同的遗传结构中使用相同的转录因子合理地创造不同类型的遗传反应的潜力。
Transcriptional interference (TI) has been shown to regulate gene expression at the DNA level via different molecular mechanisms. The obstacles present on the DNA that a transcribing RNA polymerase might encounter, for example, a DNA-bound protein or another RNA polymerase, can result in TI causing termination of transcription, thus reducing gene expression. However, the potential of TI as a new strategy to engineer complex gene expression modules has not been fully explored yet. Here we created a series of two-input genetic devices that use the presence of a roadblocking protein to control gene expression and analyzed their behaviors using both experimental and mathematical modeling approaches. We explored how multiple characteristics affect the response of genetic devices engineered to act like either AND, OR, or single input logic gates. We show that the dissociation constant of the roadblocking protein, inducer activation of promoter and operator sites, and distance between tandem promoters tune gate behavior. This work highlights the potential of rationally creating different types of genetic responses using the same transcription factors in subtly different genetic architectures.