Engineering mammalian gene activity sensor-actuator devices

工程哺乳动物基因活性传感器-致动器装置 响应环境提示生成用户定义输出的蜂窝设备史无前例 通过开发检测和纠正人类的活的设计系统为现代医学带来的机遇 病理学。这些传感器-致动器设备目前都是基于细胞表面的传感能力，大多实现 通过重新连接天然配体-受体的相互作用或进化与信号转导系统相连的非天然受体。 基于受体的传感器的性能最终取决于嵌入的信号转导机制 然而，受体系统可能不能准确地概括对生物标记物输入的生理反应。 细胞生理状态是由复杂的机制决定的，这些机制整合了与不同的 细胞外和细胞内信号的量化特征，并提供调节水平、状态和 基因表达的动力学。因此，基因表达的调节最终决定了细胞在 生理和病理过程，并不断和动态地调整，以响应环境 以及细胞内的刺激。因此，我们设想了一种新型的蜂窝设备，它可以驱动用户定义的 响应于通过实时实现的设备的生理状态的检测的生物分子程序 监测染色体基因的活性。这些基因活性传感器-致动器设备基于创新的 我们的团队最近开发了用于设计正交系统的工具，这些工具(I)将输出表达式链接到 染色体基因，从而高保真地重现复杂的哺乳动物调控过程，以及(Ii) 以高分辨率的输入动态放大信号输出，从而概括动态行为 超凡的敏感度。 生成强大的传感器-执行器设备，将基因表达签名的检测转换为用户定义的 输出，我们将探索将基因活性检测与 产出生产(目标1)，将基因活动转化为产出生产中精确调节的延迟(目标2)，自我 调整产量，以应对产量诱导的基因活性调节(目标3)。 这种方法有望在我们设计感知和响应的蜂窝设备的方式上产生一种范式转变 环境，因为它将提供一种策略来设计细胞，以感知几乎任何与 转录反应，消除了重新连接配体-受体相互作用或进化合成受体的需要 设备。这项研究的结果将产生检测基因活性高的蜂窝设备的设计规则 动态分辨率，使基于细胞的诊断和治疗技术的开发适用于各种不同的 申请。

Engineering mammalian gene activity sensors-actuator devices Cellular devices that generate user-defined outputs in response to environmental cues hold unprecedented opportunities for modern medicine through the development of living designer systems that detect and correct human pathologies. These sensor-actuator devices are currently based on cell surface sensing capabilities, mostly achieved by rewiring native ligand-receptor interactions or evolving non-native receptors linked to signal transduction systems. The performance of receptor-based sensors depends ultimately on the signal transduction mechanism embedded in the receptor system, however, and may not accurately recapitulate the physiologic response to the biomarker input. Cellular physiological states are determined by complex mechanisms that integrate signals associated with different quantitative features of extracellular and intracellular cues and provide blueprints to regulate the levels, states, and dynamics of gene expression. Regulation of gene expression thus ultimately determines cell functionality during physiological and pathological processes and is constantly and dynamically modulated to respond to environmental as well as intracellular stimuli. We thus envisioned a novel class of cellular devices that actuate user-defined biomolecular programs in response to the detection of the device’s physiological state achieved through real-time monitoring of the activity of chromosomal genes. These gene activity sensor-actuator devices are based on innovative tools recently developed by our groups for designing orthogonal systems that (i) link output expression to chromosomal genes, thereby recapitulating complex mammalian regulatory processes with high fidelity, and (ii) amplify the signal output with high resolution of the input dynamics, thereby recapitulating dynamic behaviors with superior sensitivity. To generate robust sensor-actuator devices that translate detection of gene expression signatures into user-defined outputs, we will explore the design rules of sense-and-respond mechanisms for linking detection of gene activity to output production (Aim 1), translate gene activity into precisely modulated delays in output production (Aim 2), self- adjust output production in response to output-induced modulation of gene activity (Aim 3). This approach is expected to create a paradigm shift in the way we design cellular devices that sense and respond to the environment, as it will provide a strategy to engineer cells to sense virtually any cellular process associated with a transcriptional response, eliminating the need to rewire ligand-receptor interactions or evolve synthetic receptor-based devices. Results from this study will generate design rules of cellular devices that sense gene activity with high dynamic resolution, enabling the development of cell-based diagnostics and therapeutics for a diverse range of applications.