Microcompartments for Protection and Isolation of Nanoscale DNA Computing Elements
Microcompartments for Protection and Isolation of Nanoscale DNA Computing Elements
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DOI：
10.1021/acsami.9b03143
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发表时间：
2019-03-27
影响因子：
9.5
通讯作者：
Lakin，Matthew R.
中科院分区：
文献类型：
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作者：
Fahry-Wood，Aurora;Fetrow，Madalyn Elise;Lakin，Matthew R.
分子计算元件的物理隔离通过使标准化组件能够重复使用并通过保护组件免受环境退化而具有增加系统复杂性的潜力。然而，一旦元素被划分，就需要与这些隔间进行通信的方法。我们报告了类固醇响应DNA适体在巨大的单层囊泡（GUV）内的区室化，这些囊泡可渗透类固醇输入。使用微流体平台将单分散GUV装载适体。我们证明了GUV内个别适体的靶向特异性激活，然后将两种非干扰适体加载到同一GUV中，并证明了对两种输入类固醇的所有可能组合的特异性反应。至关重要的是，GUV防止核酸酶降解DNA组分，提供了在体内部署核酸组分的潜在机制。重要的是，我们的隔间还防止互补链之间的非特异性串扰，从而提供了一种方法，用于并行执行交叉反应的分子逻辑组件。因此，我们提供了一种机制，空间组织分子计算元素，这将增加系统的模块化，允许标准化的组件被重用。
Physical isolation of molecular computing elements holds the potential for increasing system complexity by enabling the reuse of standardized components and by protecting the components from environmental degradation. However, once elements have been compartmentalized, methods for communicating into these compartments are needed. We report the compartmentalization of steroid-responsive DNA aptamers within giant unilamellar vesicles (GUVs) that are permeable to steroid inputs. Monodisperse GUVs are loaded with aptamers using a microfluidic platform. We demonstrate the target-specific activation of individual aptamers within the GUVs and then load two noninterfering aptamers into the same GUV and demonstrate specific responses to all possible combinations of the two input steroids. Crucially, GUVs prevent the degradation of DNA components by nucleases, providing a potential mechanism for deploying nucleic acid components in vivo. Importantly, our compartments also prevent nonspecific cross-talk between complementary strands, thereby providing a method for parallel execution of cross-reacting molecular logic components. Thus, we provide a mechanism for spatially organizing molecular computing elements, which will increase system modularity by allowing standardized components to be reused.